Polynucleotides, polypeptides encoded thereby, and methods of using same for increasing abiotic stress tolerance, biomass and/or yield in plants expressing same

ABSTRACT

Provided are isolated polynucleotides comprising a nucleic acid sequence at least 80% identical to SEQ ID NO:619, 617, 606, 615, 629, 1-36, 40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616, 618, 620-628, 630-638, 642, 645, 650, 651, 670, or 671. Also provided are nucleic acid constructs comprising same, isolated polypeptides encoded thereby, transgenic cells and transgenic plants comprising same and methods of using same for increasing abiotic stress tolerance, yield, biomass, growth rate, vigor, oil content, fiber yield, fiber quality, and/or nitrogen use efficiency of a plant.

RELATED APPLICATIONS

This application is a division of U.S. patent application Ser. No.13/139,729 filed on Jun. 15, 2011, which is a National Phase of PCTPatent Application No. PCT/IB2009/055962 having International filingdate of Dec. 28, 2009, which claims the benefit of priority of U.S.Provisional Patent Application Nos. 61/213,577 filed on Jun. 22, 2009;and 61/193,830 filed on Dec. 29, 2008. The contents of the aboveapplications are all incorporated herein by reference.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to isolatedpolypeptides and polynucleotides, nucleic acid constructs comprisingsame, transgenic cells comprising same, transgenic plants exogenouslyexpressing same and more particularly, but not exclusively, to methodsof using same for increasing abiotic stress tolerance, growth rate,biomass, vigor, yield (e.g., seed yield, oil yield), oil content, fiberyield, fiber quality and/or fertilizer use efficiency (e.g., nitrogenuse efficiency) of a plant.

Abiotic stress (ABS; also referred to as “environmental stress”)conditions such as salinity, drought, flood, suboptimal temperature andtoxic chemical pollution, cause substantial damage to agriculturalplants. Most plants have evolved strategies to protect themselvesagainst these conditions. However, if the severity and duration of thestress conditions are too great, the effects on plant development,growth and yield of most crop plants are profound. Furthermore, most ofthe crop plants are highly susceptible to abiotic stress and thusnecessitate optimal growth conditions for commercial crop yields.Continuous exposure to stress causes major alterations in the plantmetabolism which ultimately leads to cell death and consequently yieldlosses.

The global shortage of water supply is one of the most severeagricultural problems affecting plant growth and crop yield and effortsare made to mitigate the harmful effects of desertification andsalinization of the world's arable land. Water deficit is a commoncomponent of many plant stresses and occurs in plant cells when thewhole plant transpiration rate exceeds the water uptake. In addition todrought, other stresses, such as salinity and low temperature, producecellular dehydration.

Drought is a gradual phenomenon, which involves periods of abnormallydry weather that persists long enough to produce serious hydrologicimbalances such as crop damage and water supply shortage. In severecases, drought can last many years and results in devastating effects onagriculture and water supplies. Furthermore, drought is associated withincrease susceptibility to various diseases.

For most crop plants, the land regions of the world are too arid. Inaddition, overuse of available water results in increased loss ofagriculturally-usable land (desertification), and increase of saltaccumulation in soils adds to the loss of available water in soils.

Salinity, high salt levels, affects one in five hectares of irrigatedland. This condition is only expected to worsen, further reducing theavailability of arable land and crop production, since none of the topfive food crops, i.e., wheat, corn, rice, potatoes, and soybean, cantolerate excessive salt. Detrimental effects of salt on plants resultfrom both water deficit which leads to osmotic stress (similar todrought stress) and the effect of excess sodium ions on criticalbiochemical processes. As with freezing and drought, high salt causeswater deficit; and the presence of high salt makes it difficult forplant roots to extract water from their environment. Soil salinity isthus one of the more important variables that determine whether a plantmay thrive. In many parts of the world, sizable land areas areuncultivable due to naturally high soil salinity. Thus, salination ofsoils that are used for agricultural production is a significant andincreasing problem in regions that rely heavily on agriculture, and isworsen by over-utilization, over-fertilization and water shortage,typically caused by climatic change and the demands of increasingpopulation. Salt tolerance is of particular importance early in aplant's lifecycle, since evaporation from the soil surface causes upwardwater movement, and salt accumulates in the upper soil layer where theseeds are placed. On the other hand, germination normally takes place ata salt concentration which is higher than the mean salt level in thewhole soil profile.

Germination of many crops is sensitive to temperature. A gene that wouldenhance germination in hot conditions would be useful for crops that areplanted late in the season or in hot climates. In addition, seedlingsand mature plants that are exposed to excess heat may experience heatshock, which may arise in various organs, including leaves andparticularly fruit, when transpiration is insufficient to overcome heatstress. Heat also damages cellular structures, including organelles andcyto skeleton, and impairs membrane function. Heat shock may produce adecrease in overall protein synthesis, accompanied by expression of heatshock proteins, e.g., chaperones, which are involved in refoldingproteins denatured by heat.

Heat stress often accompanies conditions of low water availability. Heatitself is seen as an interacting stress and adds to the detrimentaleffects caused by water deficit conditions. Water evaporation increasesalong with the rise in daytime temperatures and can result in hightranspiration rates and low plant water potentials. High-temperaturedamage to pollen almost always occurs in conjunction with droughtstress, and rarely occurs under well-watered conditions. Combined stresscan alter plant metabolism in various ways; therefore understanding theinteraction between different stresses may be important for thedevelopment of strategies to enhance stress tolerance by geneticmanipulation.

Excessive chilling conditions, e.g., low, but above freezing,temperatures affect crops of tropical origins, such as soybean, rice,maize, and cotton. Typical chilling damage includes wilting, necrosis,chlorosis or leakage of ions from cell membranes. The underlyingmechanisms of chilling sensitivity are not completely understood yet,but probably involve the level of membrane saturation and otherphysiological deficiencies. For example, photoinhibition ofphotosynthesis (disruption of photosynthesis due to high lightintensities) often occurs under clear atmospheric conditions subsequentto cold late summer/autumn nights. In addition, chilling may lead toyield losses and lower product quality through the delayed ripening ofmaize.

Salt and drought stress signal transduction consist of ionic and osmotichomeostasis signaling pathways. The ionic aspect of salt stress issignaled via the SOS pathway where a calcium-responsive SOS3-SOS2protein kinase complex controls the expression and activity of iontransporters such as SOS1. The osmotic component of salt stress involvescomplex plant reactions that overlap with drought and/or cold stressresponses.

Common aspects of drought, cold and salt stress response [Reviewed inXiong and Zhu (2002) Plant Cell Environ. 25: 131-139] include: (a)transient changes in the cytoplasmic calcium levels early in thesignaling event; (b) signal transduction via mitogen-activated and/orcalcium dependent protein kinases (CDPKs) and protein phosphatases; (c)increases in abscisic acid levels in response to stress triggering asubset of responses; (d) inositol phosphates as signal molecules (atleast for a subset of the stress responsive transcriptional changes; (e)activation of phospholipases which in turn generates a diverse array ofsecond messenger molecules, some of which might regulate the activity ofstress responsive kinases; (f) induction of late embryogenesis abundant(LEA) type genes including the CRT/DRE responsive COR/RD genes; (g)increased levels of antioxidants and compatible osmolytes such asproline and soluble sugars; and (h) accumulation of reactive oxygenspecies such as superoxide, hydrogen peroxide, and hydroxyl radicals.Abscisic acid biosynthesis is regulated by osmotic stress at multiplesteps. Both ABA-dependent and -independent osmotic stress signalingfirst modify constitutively expressed transcription factors, leading tothe expression of early response transcriptional activators, which thenactivate downstream stress tolerance effector genes.

Several genes which increase tolerance to cold or salt stress can alsoimprove drought stress protection, these include for example, thetranscription factor AtCBF/DREB1, OsCDPK7 (Saijo et al. 2000, Plant J.23: 319-327) or AVP1 (a vacuolar pyrophosphatase-proton pump, Gaxiola etal. 2001, Proc. Natl. Acad. Sci. USA 98: 11444-11449).

Developing stress-tolerant plants is a strategy that has the potentialto solve or mediate at least some of these problems. However,traditional plant breeding strategies used to develop new lines ofplants that exhibit tolerance to ABS are relatively inefficient sincethey are tedious, time consuming and of unpredictable outcome.Furthermore, limited germplasm resources for stress tolerance andincompatibility in crosses between distantly related plant speciesrepresent significant problems encountered in conventional breeding.Additionally, the cellular processes leading to ABS tolerance arecomplex in nature and involve multiple mechanisms of cellular adaptationand numerous metabolic pathways.

Genetic engineering efforts, aimed at conferring abiotic stresstolerance to transgenic crops, have been described in variouspublications [Apse and Blumwald (Curr Opin Biotechnol. 13:146-150,2002), Quesada et al. (Plant Physiol. 130:951-963, 2002), Holmström etal. (Nature 379: 683-684, 1996), Xu et al. (Plant Physiol 110: 249-257,1996), Pilon-Smits and Ebskamp (Plant Physiol 107: 125-130, 1995) andTarczynski et al. (Science 259: 508-510, 1993)].

Various patents and patent applications disclose genes and proteinswhich can be used for increasing tolerance of plants to abioticstresses. These include for example, U.S. Pat. Nos. 5,296,462 and5,356,816 (for increasing tolerance to cold stress); U.S. Pat. No.6,670,528 (for increasing ABST); U.S. Pat. No. 6,720,477 (for increasingABST); U.S. application Ser. Nos. 09/938,842 and 10/342,224 (forincreasing ABST); U.S. application Ser. No. 10/231,035 (for increasingABST); WO2004/104162 (for increasing ABST and biomass); WO2007/020638(for increasing ABST, biomass, vigor and/or yield); WO2007/049275 (forincreasing ABST, biomass, vigor and/or yield).

Suboptimal nutrient (macro and micro nutrient) affect plant growth anddevelopment through the whole plant life cycle. One of the essentialmacronutrients for the plant is Nitrogen. Nitrogen is responsible forbiosynthesis of amino acids and nucleic acids, prosthetic groups, planthormones, plant chemical defenses, and the like. Nitrogen is often therate-limiting element in plant growth and all field crops have afundamental dependence on inorganic nitrogenous fertilizer. Sincefertilizer is rapidly depleted from most soil types, it must be suppliedto growing crops two or three times during the growing season.Additional important macronutrients are Phosphorous (P) and Potassium(K), which have a direct correlation to yield and general planttolerance.

Vegetable or seed oils are the major source of energy and nutrition inhuman and animal diet. They are also used for the production ofindustrial products, such as paints, inks and lubricants. In addition,plant oils represent renewable sources of long-chain hydrocarbons whichcan be used as fuel. Since the currently used fossil fuels are finiteresources and are gradually being depleted, fast growing biomass cropsmay be used as alternative fuels or for energy feedstocks and may reducethe dependence on fossil energy supplies. However, the major bottleneckfor increasing consumption of plant oils as bio-fuel is the oil price,which is still higher than fossil fuel. In addition, the production rateof plant oil is limited by the availability of agricultural land andwater. Thus, increasing plant oil yields from the same growing area caneffectively overcome the shortage in production space and can decreasevegetable oil prices at the same time.

Studies aiming at increasing plant oil yields focus on theidentification of genes involved in oil metabolism as well as in genescapable of increasing plant and seed yields in transgenic plants. Genesknown to be involved in increasing plant oil yields include thoseparticipating in fatty acid synthesis or sequestering such as desaturase[e.g., DELTA6, DELTA12 or acyl-ACP (Ssi2; Arabidopsis InformationResource (TAIR; Hypertext Transfer Protocol://World Wide Web (dot)arabidopsis (dot) org/), TAIR No. AT2G43710)], OleosinA (TAIR No.AT3G01570) or FAD3 (TAIR No. AT2G29980), and various transcriptionfactors and activators such as Lec1 [TAIR No. AT1G21970, Lotan et al.1998. Cell. 26; 93(7):1195-205], Lec2 [TAIR No. AT1G28300, SantosMendoza et al. 2005, FEBS Lett. 579(20:4666-70], Fus3 (TAIR No.AT3G26790), ABI3 [TAIR No. AT3G24650, Lara et al. 2003. J Biol. Chem.278(23): 21003-11] and Wri1 [TAIR No. AT3G54320, Cernac and Benning,2004. Plant J. 40(4): 575-85].

Genetic engineering efforts aiming at increasing oil content in plants(e.g., in seeds) include upregulating endoplasmic reticulum (FAD3) andplastidal (FAD7) fatty acid desaturases in potato (Zabrouskov V., etal., 2002; Physiol Plant. 116:172-185); over-expressing the GmDof4 andGmDof11 transcription factors (Wang H W et al., 2007; Plant J.52:716-29); over-expressing a yeast glycerol-3-phosphate dehydrogenaseunder the control of a seed-specific promoter (Vigeolas H, et al. 2007,Plant Biotechnol J. 5:431-41; U.S. Pat. Appl. No. 20060168684); usingArabidopsis FAE1 and yeast SLC1-1 genes for improvements in erucic acidand oil content in rapeseed (Katavic V, et al., 2000, Biochem Soc Trans.28:935-7).

Various patent applications disclose genes and proteins which canincrease oil content in plants. These include for example, U.S. Pat.Appl. No. 20080076179 (lipid metabolism protein); U.S. Pat. Appl. No.20060206961 (the Ypr140w polypeptide); U.S. Pat. Appl. No. 20060174373[triacylglycerols synthesis enhancing protein (TEP)]; U.S. Pat. Appl.Nos. 20070169219, 20070006345, 20070006346 and 20060195943 (disclosetransgenic plants with improved nitrogen use efficiency which can beused for the conversion into fuel or chemical feedstocks); WO2008/122980(polynucleotides for increasing oil content, growth rate, biomass, yieldand/or vigor of a plant).

Cotton and cotton by-products provide raw materials that are used toproduce a wealth of consumer-based products in addition to textilesincluding cotton foodstuffs, livestock feed, fertilizer and paper. Theproduction, marketing, consumption and trade of cotton-based productsgenerate an excess of $100 billion annually in the U.S. alone, makingcotton the number one value-added crop.

Even though 90% of cotton's value as a crop resides in the fiber (lint),yield and fiber quality has declined due to general erosion in geneticdiversity of cotton varieties, and an increased vulnerability of thecrop to environmental conditions.

There are many varieties of cotton plant, from which cotton fibers witha range of characteristics can be obtained and used for variousapplications. Cotton fibers may be characterized according to a varietyof properties, some of which are considered highly desirable within thetextile industry for the production of increasingly high qualityproducts and optimal exploitation of modem spinning technologies.Commercially desirable properties include length, length uniformity,fineness, maturity ratio, decreased fuzz fiber production, micronaire,bundle strength, and single fiber strength. Much effort has been putinto the improvement of the characteristics of cotton fibers mainlyfocusing on fiber length and fiber fineness. In particular, there is agreat demand for cotton fibers of specific lengths.

A cotton fiber is composed of a single cell that has differentiated froman epidermal cell of the seed coat, developing through four stages,i.e., initiation, elongation, secondary cell wall thickening andmaturation stages. More specifically, the elongation of a cotton fibercommences in the epidermal cell of the ovule immediately followingflowering, after which the cotton fiber rapidly elongates forapproximately 21 days. Fiber elongation is then terminated, and asecondary cell wall is formed and grown through maturation to become amature cotton fiber.

Several candidate genes which are associated with the elongation,formation, quality and yield of cotton fibers were disclosed in variouspatent applications such as U.S. Pat. No. 5,880,100 and U.S. patentapplication Ser. Nos. 08/580,545, 08/867,484 and Ser. No. 09/262,653(describing genes involved in cotton fiber elongation stage); WO0245485(improving fiber quality by modulating sucrose synthase); U.S. Pat. No.6,472,588 and WO0117333 (increasing fiber quality by transformation witha DNA encoding sucrose phosphate synthase); WO9508914 (using afiber-specific promoter and a coding sequence encoding cottonperoxidase); WO9626639 (using an ovary specific promoter sequence toexpress plant growth modifying hormones in cotton ovule tissue, foraltering fiber quality characteristics such as fiber dimension andstrength); U.S. Pat. No. 5,981,834, U.S. Pat. No. 5,597,718, U.S. Pat.No. 5,620,882, U.S. Pat. No. 5,521,708 and U.S. Pat. No. 5,495,070(coding sequences to alter the fiber characteristics of transgenic fiberproducing plants); U.S. patent applications U.S. 2002049999 and U.S.2003074697 (expressing a gene coding for endoxyloglucan transferase,catalase or peroxidase for improving cotton fiber characteristics); WO01/40250 (improving cotton fiber quality by modulating transcriptionfactor gene expression); WO 96/40924 (a cotton fiber transcriptionalinitiation regulatory region associated which is expressed in cottonfiber); EP0834566 (a gene which controls the fiber formation mechanismin cotton plant); WO2005/121364 (improving cotton fiber quality bymodulating gene expression); WO2008/075364 (improving fiber quality,yield/biomass/vigor and/or abiotic stress tolerance of plants).

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence at least 80% identical to SEQ ID NO:619, 617, 606,615, 629, 1-36, 40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358,605, 607-614, 616, 618, 620-628, 630-638, 642, 645, 650, 651, 670, or671, thereby increasing the abiotic stress tolerance, yield, biomass,growth rate, vigor, oil content, fiber yield, fiber quality, and/ornitrogen use efficiency of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising thenucleic acid sequence selected from the group consisting of SEQ IDNOs:619, 617, 606, 615, 629, 1-49, 51-59, 113-343, 345-351, 353-358,605, 607-614, 616, 618, 620-628, 630-638, 641, 642, 644, 644-646,648-651, 670, and 671, thereby increasing the abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,nitrogen use efficiency, fiber yield and/or fiber quality of a plant,comprising expressing within the plant an exogenous polynucleotidecomprising a nucleic acid sequence at least 80% identical to SEQ ID NO:352, 639, 640, or 643, thereby increasing the abiotic stress tolerance,nitrogen use efficiency, fiber yield and/or fiber quality of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing nitrogen use efficiency, seedyield and/or oil content of a plant, comprising expressing within theplant an exogenous polynucleotide comprising a nucleic acid sequence atleast 80% identical to SEQ ID NO: 50, 645, or 647, thereby increasingthe nitrogen use efficiency, seed yield and/or oil content of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing seed yield, fiber yield and/orfiber quality of a plant, comprising expressing within the plant anexogenous polynucleotide comprising a nucleic acid sequence at least 80%identical to SEQ ID NO:344, thereby increasing the seed yield, fiberyield and/or fiber quality of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence encoding a polypeptide at least 80% identical toSEQ ID NO:75, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-95, 108-109,112, 359-589, 602-604, 653-660, 665, 668, or 672, thereby increasing theabiotic stress tolerance, yield, biomass, growth rate, vigor, oilcontent, fiber yield, fiber quality, and/or nitrogen use efficiency ofthe plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence encoding a polypeptide selected from the groupconsisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72, 74, 76-85,87-98, 100-109, 111, 112, 359-589, 591-597, 600-604, 653-662, 664,666-669, and 672, thereby increasing the abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing abiotic stress tolerance,nitrogen use efficiency, fiber yield and/or fiber quality of a plant,comprising expressing within the plant an exogenous polynucleotidecomprising a nucleic acid sequence encoding a polypeptide at least 80%identical to SEQ ID NO:99 or 598, thereby increasing the abiotic stresstolerance, nitrogen use efficiency, fiber yield and/or fiber quality ofthe plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing nitrogen use efficiency, seedyield and/or oil content of a plant, comprising expressing within theplant an exogenous polynucleotide comprising a nucleic acid sequenceencoding a polypeptide at least 80% identical to SEQ ID NO:599 or 663,thereby increasing the nitrogen use efficiency, seed yield and/or oilcontent of the plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing nitrogen use efficiency,abiotic stress tolerance, seed yield and/or oil content of a plant,comprising expressing within the plant an exogenous polynucleotidecomprising a nucleic acid sequence encoding a polypeptide at least 80%identical to SEQ ID NO:110 or 665, thereby increasing the nitrogen useefficiency, abiotic stress tolerance, seed yield and/or oil content ofthe plant.

According to an aspect of some embodiments of the present inventionthere is provided a method of increasing seed yield, fiber yield and/orfiber quality of a plant, comprising expressing within the plant anexogenous polynucleotide comprising a nucleic acid sequence encoding apolypeptide at least 80% identical to SEQ ID NO:590, thereby increasingthe seed yield, fiber yield and/or fiber quality of the plant.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polynucleotide comprising a nucleic acidsequence at least 80% identical to SEQ ID NO:619, 617, 606, 615, 629,1-36, 40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614,616, 618, 620-628, 630-638, 642, 645, 650-651, 670, or 671, wherein saidnucleic acid sequence is capable of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polynucleotide comprising the nucleic acidsequence selected from the group consisting of SEQ ID NOs:619, 617, 606,615, 629, 1-49, 51-59, 113-343, 345-351, 353-358, 605, 607-614, 616,618, 620-628, 630-638, 641, 642, 644, 644-646, 648-651, 670, and 671.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polynucleotide comprising a nucleic acidsequence encoding a polypeptide which comprises an amino acid sequenceat least 80% homologous to the amino acid sequence set forth in SEQ IDNO: 75, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-95, 108-109, 112,359-589, 602-604, 653-660, 665, 668, or 672, wherein said amino acidsequence is capable of increasing abiotic stress tolerance, yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of a plant.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polynucleotide comprising a nucleic acidsequence encoding a polypeptide which comprises the amino acid sequenceselected from the group consisting of SEQ ID NOs:75, 73, 652, 71, 86,60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589, 591-597,600-604, 653-662, 664, 666-669, and 672.

According to an aspect of some embodiments of the present inventionthere is provided a nucleic acid construct comprising the isolatedpolynucleotide of claim 12, 13, 14 or 15, and a promoter for directingtranscription of said nucleic acid sequence in a host cell.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polypeptide comprising an amino acidsequence at least 80% homologous to SEQ ID NO:75, 73, 652, 71, 86,60-70, 72, 74, 76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660,665, 668, or 672, wherein said amino acid sequence is capable ofincreasing abiotic stress tolerance, yield, biomass, growth rate, vigor,oil content, fiber yield, fiber quality, and/or nitrogen use efficiencyof a plant.

According to an aspect of some embodiments of the present inventionthere is provided an isolated polypeptide comprising the amino acidsequence selected from the group consisting of SEQ ID NOs:75, 73, 652,71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589,591-597, 600-604, 653-662, 664, 666-669, and 672

According to an aspect of some embodiments of the present inventionthere is provided a plant cell exogenously expressing the polynucleotideof claim 12, 13, 14 or 15, or the nucleic acid construct of claim 16.

According to an aspect of some embodiments of the present inventionthere is provided a plant cell exogenously expressing the polypeptide ofclaim 17 or 18.

According to some embodiments of the invention, the nucleic acidsequence is as set forth in SEQ ID NO:619, 617, 606, 615, 629, 1-36, 40,41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616, 618,620-628, 630-638, 642, 645, 650, 651, 670, or 671.

According to some embodiments of the invention, the polynucleotideconsists of the nucleic acid sequence selected from the group consistingof SEQ ID NOs:619, 617, 606, 615, 629, 1-36, 40, 41, 43-45, 49, 52-56,58, 113-343, 351, 354-358, 605, 607-614, 616, 618, 620-628, 630-638,642, 645, 650, 651, 670, and 671.

According to some embodiments of the invention, the nucleic acidsequence encodes an amino acid sequence at least 80% homologous to SEQID NO:75, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-95, 108-109, 112,359-589, 602-604, 653-660, 665, 668, or 672.

According to some embodiments of the invention, the nucleic acidsequence encodes the amino acid sequence selected from the groupconsisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72, 74, 76-85,87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, and 672.

According to some embodiments of the invention, the plant cell formspart of a plant.

According to some embodiments of the invention, the method furthercomprising growing the plant expressing said exogenous polynucleotideunder the abiotic stress.

According to some embodiments of the invention, the abiotic stress isselected from the group consisting of salinity, drought, waterdeprivation, flood, etiolation, low temperature, high temperature, heavymetal toxicity, anaerobiosis, nutrient deficiency, nutrient excess,atmospheric pollution and UV irradiation.

According to some embodiments of the invention, the yield comprises seedyield or oil yield.

Unless otherwise defined, all technical and/or scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which the invention pertains. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of embodiments of the invention, exemplarymethods and/or materials are described below. In case of conflict, thepatent specification, including definitions, will control. In addition,the materials, methods, and examples are illustrative only and are notintended to be necessarily limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

Some embodiments of the invention are herein described, by way ofexample only, with reference to the accompanying drawings. With specificreference now to the drawings in detail, it is stressed that theparticulars shown are by way of example and for purposes of illustrativediscussion of embodiments of the invention. In this regard, thedescription taken with the drawings makes apparent to those skilled inthe art how embodiments of the invention may be practiced.

In the drawings:

FIG. 1 is a schematic illustration of the pGI binary plasmid used forexpressing the isolated polynucleotide sequences of some embodiments ofthe invention. RB—T-DNA right border; LB—T-DNA left border; H—HindIIIrestriction enzyme; X—XbaI restriction enzyme; B—BamHI restrictionenzyme; S—SaiI restriction enzyme; Sm—SmaI restriction enzyme; R-I—EcoRIrestriction enzyme; Sc—SacI/SstI/Ecl136II; (numbers)—Length inbase-pairs; NOS pro=nopaline synthase promoter; NPT-II=neomycinphosphotransferase gene; NOS ter=nopaline synthase terminator; Poly-Asignal (polyadenylation signal); GUSintron—the GUS reporter gene (codingsequence and intron) The isolated polynucleotide sequences of theinvention were cloned into the vector while replacing the GUSintronreporter gene

FIG. 2 is a schematic illustration of the modified pGI binary plasmidused for expressing the isolated polynucleotide sequences of theinvention. RB—T-DNA right border; LB—T-DNA left border; MCS—Multiplecloning site; RE—any restriction enzyme; (numbers)—Length in base-pairs;NOS pro=nopaline synthase promoter; NPT-II=neomycin phosphotransferasegene; NOS ter=nopaline synthase terminator; Poly-A signal(polyadenylation signal); GUSintron—the GUS reporter gene (codingsequence and intron) The isolated polynucleotide sequences of theinvention were cloned into the vector while replacing the GUSintronreporter gene.

FIGS. 3A-F are images depicting visualization of root development oftransgenic plants exogenously expressing the polynucleotide of someembodiments of the invention when grown in transparent agar plates undernormal (FIGS. 3A-B), osmotic stress (15% PEG; FIGS. 3C-D) ornitrogen-limiting (FIGS. 3E-F) conditions. The different transgenes weregrown in transparent agar plates for 17 days (7 days nursery and 10 daysafter transplanting). The plates were photographed every 3-4 daysstarting at day 1 after transplanting. FIG. 3A—An image of a photographof plants taken following 10 after transplanting days on agar plateswhen grown under normal (standard) conditions. FIG. 3B—An image of rootanalysis of the plants shown in FIG. 3A in which the lengths of theroots measured are represented by arrows. FIG. 3C—An image of aphotograph of plants taken following 10 days after transplanting on agarplates, grown under high osmotic (PEG 15%) conditions. FIG. 3D—An imageof root analysis of the plants shown in FIG. 3C in which the lengths ofthe roots measured are represented by arrows. FIG. 3E—An image of aphotograph of plants taken following 10 days after transplanting on agarplates, grown under low nitrogen conditions. FIG. 3F—An image of rootanalysis of the plants shown in FIG. 3E in which the lengths of theroots measured are represented by arrows.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention relates to polypeptides, polynucleotides, nucleicacid constructs and methods of increasing abiotic stress tolerance,fertilizer use efficiency (e.g., nitrogen use efficiency), growth,biomass, fiber development or quality, vigor and/or yield of a plant.

Before explaining at least one embodiment of the invention in detail, itis to be understood that the invention is not necessarily limited in itsapplication to the details set forth in the following description orexemplified by the Examples. The invention is capable of otherembodiments or of being practiced or carried out in various ways.

The present inventors have identified novel polypeptides andpolynucleotides which can be used to increase abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant.

Thus, as shown in the Examples section which follows, the presentinventors have utilized bioinformatics tools to identify polynucleotideswhich increase abiotic stress tolerance (ABST), fertilizer useefficiency [e.g., nitrogen use efficiency (NUE)], yield (e.g., seedyield, oil yield, oil content), growth rate, biomass, vigor and/or of aplant. Genes which affect the trait-of-interest were identified usingdigital expression profiles in specific tissues and conditions such asexpression in roots; expression under stress conditions such as droughtstress, ultraviolet (UV) irradiation, cold stress, heat stress, nutrientdeficiency, stress hormones [for example as abscisic acid (ABA) andethylene] etiolation conditions, salinity stress, waterlogging; and/orexpression during plant development (Tables 1-5; Example 1 of theExamples section which follows; polynucleotide SEQ ID NOs:1-59 and 638;polypeptide SEQ ID NOs:60-112). Homologous polypeptides andpolynucleotides having the same function were also identified (Table 6,Example 2 of the Examples section which follows; polynucleotide SEQ IDNOs:113-358; polypeptide SEQ ID NOs:359-604). The identifiedpolynucleotides were cloned into binary vectors [Tables 7-10; Example 3;SEQ ID NOs:605-637, 639-651, 670, 671 (polynucleotides); SEQ ID NOs: 60,63-73, 75, 77, 81-83, 86, 87, 90, 92, 93, 94, 95, 96, 99, 100, 101, 102,652-669, 672 (polypeptides)], transformed into agrobacterium cells(Example 4), and further into Arabidopsis plants (Example 5). Transgenicplants over-expressing the identified polynucleotides were found toexhibit increased biomass [e.g., fresh and dry weight; leaf area andgrowth rate, rosette area, rosette diameter and growth rate of rosettearea and diameter, plot coverage, leaf number], growth rate, yield(e.g., seed yield and weight), harvest index, roots growth (e.g., rootlength, root coverage, growth rate of root length and/or coverage), oilyield, oil percentage in seeds, weight of 1000 seeds (Tables 11-62;Examples 6, 7, 8, 9, and 11 of the Examples section which follows) undernormal or limiting conditions (e.g., abiotic stress, nitrogen limitingconditions). Altogether, these results suggest the use of the novelpolynucleotides and polypeptides of the invention for increasing abioticstress tolerance, yield, biomass, growth rate, vigor, oil content, fiberyield, fiber quality, and/or nitrogen use efficiency of a plant.

Thus, according to an aspect of some embodiments of the invention, thereis provided method of increasing abiotic stress tolerance, yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of a plant, the method comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence at least 80% identical to SEQ ID NO: 619, 617,606, 615, 629, 1-36, 40, 41, 43-45, 49, 52-56, 58, 113-343, 351,354-358, 605, 607-614, 616, 618, 620-628, 630-638, 642, 645, 650, 651,670 or 671, thereby increasing the abiotic stress tolerance, yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of the plant.

As used herein the phrase “plant yield” refers to the amount (e.g., asdetermined by weight or size) or quantity (numbers) of tissues or organsproduced per plant or per growing season. Hence increased yield couldaffect the economic benefit one can obtain from the plant in a certaingrowing area and/or growing time.

It should be noted that a plant yield can be affected by variousparameters including, but not limited to, plant biomass; plant vigor;growth rate; seed yield; seed or grain quantity; seed or grain quality;oil yield; content of oil, starch and/or protein in harvested organs(e.g., seeds or vegetative parts of the plant); number of flowers(florets) per panicle (expressed as a ratio of number of filled seedsover number of primary panicles); harvest index; number of plants grownper area; number and size of harvested organs per plant and per area;number of plants per growing area (density); number of harvested organsin field; total leaf area; carbon assimilation and carbon partitioning(the distribution/allocation of carbon within the plant); resistance toshade; number of harvestable organs (e.g. seeds), seeds per pod, weightper seed; and modified architecture [such as increase stalk diameter,thickness or improvement of physical properties (e.g. elasticity)].

As used herein the phrase “seed yield” refers to the number or weight ofthe seeds per plant, seeds per pod, or per growing area or to the weightof a single seed, or to the oil extracted per seed. Hence seed yield canbe affected by seed dimensions (e.g., length, width, perimeter, areaand/or volume), number of (filled) seeds and seed filling rate and byseed oil content. Hence increase seed yield per plant could affect theeconomic benefit one can obtain from the plant in a certain growing areaand/or growing time; and increase seed yield per growing area could beachieved by increasing seed yield per plant, and/or by increasing numberof plants grown on the same given area.

The term “seed” (also referred to as “grain” or “kernel”) as used hereinrefers to a small embryonic plant enclosed in a covering called the seedcoat (usually with some stored food), the product of the ripened ovuleof gymnosperm and angiosperm plants which occurs after fertilization andsome growth within the mother plant.

The phrase “oil content” as used herein refers to the amount of lipidsin a given plant organ, either the seeds (seed oil content) or thevegetative portion of the plant (vegetative oil content) and istypically expressed as percentage of dry weight (10% humidity of seeds)or wet weight (for vegetative portion).

It should be noted that oil content is affected by intrinsic oilproduction of a tissue (e.g., seed, vegetative portion), as well as themass or size of the oil-producing tissue per plant or per growth period.

In one embodiment, increase in oil content of the plant can be achievedby increasing the size/mass of a plant's tissue(s) which comprise oilper growth period. Thus, increased oil content of a plant can beachieved by increasing the yield, growth rate, biomass and vigor of theplant.

As used herein the phrase “plant biomass” refers to the amount (e.g.,measured in grams of air-dry tissue) of a tissue produced from the plantin a growing season, which could also determine or affect the plantyield or the yield per growing area. An increase in plant biomass can bein the whole plant or in parts thereof such as aboveground (harvestable)parts, vegetative biomass, roots and seeds.

As used herein the phrase “growth rate” refers to the increase in plantorgan/tissue size per time (can be measured in cm² per day).

As used herein the phrase “plant vigor” refers to the amount (measuredby weight) of tissue produced by the plant in a given time. Henceincreased vigor could determine or affect the plant yield or the yieldper growing time or growing area. In addition, early vigor (seed and/orseedling) results in improved field stand.

It should be noted that a plant yield can be determined under stress(e.g., abiotic stress, nitrogen-limiting conditions) and/or non-stress(normal) conditions.

As used herein, the phrase “non-stress conditions” refers to the growthconditions (e.g., water, temperature, light-dark cycles, humidity, saltconcentration, fertilizer concentration in soil, nutrient supply such asnitrogen, phosphorous and/or potassium), that do not significantly gobeyond the everyday climatic and other abiotic conditions that plantsmay encounter, and which allow optimal growth, metabolism, reproductionand/or viability of a plant at any stage in its life cycle (e.g., in acrop plant from seed to a mature plant and back to seed again). Personsskilled in the art are aware of normal soil conditions and climaticconditions for a given plant in a given geographic location. It shouldbe noted that while the non-stress conditions may include some mildvariations from the optimal conditions (which vary from one type/speciesof a plant to another), such variations do not cause the plant to ceasegrowing without the capacity to resume growth.

The phrase “abiotic stress” as used herein refers to any adverse effecton metabolism, growth, reproduction and/or viability of a plant.Accordingly, abiotic stress can be induced by suboptimal environmentalgrowth conditions such as, for example, salinity, water deprivation,flooding, freezing, low or high temperature, heavy metal toxicity,anaerobiosis, nutrient deficiency, atmospheric pollution or UVirradiation. The implications of abiotic stress are discussed in theBackground section.

The phrase “abiotic stress tolerance” as used herein refers to theability of a plant to endure an abiotic stress without suffering asubstantial alteration in metabolism, growth, productivity and/orviability.

As used herein the phrase “water use efficiency (WUE)” refers to thelevel of organic matter produced per unit of water consumed by theplant, i.e., the dry weight of a plant in relation to the plant's wateruse, e.g., the biomass produced per unit transpiration.

As used herein the phrase “fertilizer use efficiency” refers to themetabolic process(es) which lead to an increase in the plant's yield,biomass, vigor, and growth rate per fertilizer unit applied. Themetabolic process can be the uptake, spread, absorbent, accumulation,relocation (within the plant) and use of one or more of the minerals andorganic moieties absorbed by the plant, such as nitrogen, phosphatesand/or potassium.

As used herein the phrase “fertilizer-limiting conditions” refers togrowth conditions which include a level (e.g., concentration) of afertilizer applied which is below the level needed for normal plantmetabolism, growth, reproduction and/or viability.

As used herein the phrase “nitrogen use efficiency (NUE)” refers to themetabolic process(es) which lead to an increase in the plant's yield,biomass, vigor, and growth rate per nitrogen unit applied. The metabolicprocess can be the uptake, spread, absorbent, accumulation, relocation(within the plant) and use of nitrogen absorbed by the plant.

As used herein the phrase “nitrogen-limiting conditions” refers togrowth conditions which include a level (e.g., concentration) ofnitrogen (e.g., ammonium or nitrate) applied which is below the levelneeded for normal plant metabolism, growth, reproduction and/orviability.

Improved plant NUE and FUE is translated in the field into eitherharvesting similar quantities of yield, while implementing lessfertilizers, or increased yields gained by implementing the same levelsof fertilizers. Thus, improved NUE or FUE has a direct effect on plantyield in the field. Thus, the polynucleotides and polypeptides of someembodiments of the invention positively affect plant yield, seed yield,and plant biomass. In addition, the benefit of improved plant NUE willcertainly improve crop quality and biochemical constituents of the seedsuch as protein yield and oil yield.

It should be noted that improved ABST will confer plants with improvedvigor also under non-stress conditions, resulting in crops havingimproved biomass and/or yield e.g., elongated fibers for the cottonindustry, higher oil content.

As used herein the term “increasing” refers to at least about 2%, atleast about 3%, at least about 4%, at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 30%, atleast about 40%, at least about 50%, at least about 60%, at least about70%, at least about 80%, increase in abiotic stress tolerance, yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of a plant as compared to a native plant[i.e., a plant not modified with the biomolecules (polynucleotide orpolypeptides) of the invention, e.g., a non-transformed plant of thesame species which is grown under the same growth conditions).

The phrase “expressing within the plant an exogenous polynucleotide” asused herein refers to upregulating the expression level of an exogenouspolynucleotide within the plant by introducing the exogenouspolynucleotide into a plant cell or plant and expressing by recombinantmeans, as further described herein below.

As used herein “expressing” refers to expression at the mRNA andoptionally polypeptide level.

As used herein, the phrase “exogenous polynucleotide” refers to aheterologous nucleic acid sequence which may not be naturally expressedwithin the plant or which overexpression in the plant is desired. Theexogenous polynucleotide may be introduced into the plant in a stable ortransient manner, so as to produce a ribonucleic acid (RNA) moleculeand/or a polypeptide molecule. It should be noted that the exogenouspolynucleotide may comprise a nucleic acid sequence which is identicalor partially homologous to an endogenous nucleic acid sequence of theplant.

The term “endogenous” as used herein refers to any polynucleotide orpolypeptide which is present and/or naturally expressed within a plantor a cell thereof.

According to some embodiments of the invention the exogenouspolynucleotide comprises a nucleic acid sequence which is at least about80%, at least about 81%, at least about 82%, at least about 83%, atleast about 84%, at least about 85%, at least about 86%, at least about87%, at least about 88%, at least about 89%, at least about 90%, atleast about 91%, at least about 92%, at least about 93%, at least about93%, at least about 94%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, e.g., 100%identical to the nucleic acid sequence selected from the groupconsisting of SEQ ID NOs: 619, 617, 606, 615, 629, 1-36, 40, 41, 43-45,49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616, 618, 620-628,630-638, 642, 645, 650, 651, 670, and 671.

Identity (e.g., percent homology) can be determined using any homologycomparison software, including for example, the BlastN software of theNational Center of Biotechnology Information (NCBI) such as by usingdefault parameters.

According to some embodiments of the invention the exogenouspolynucleotide is at least about 80%, at least about 81%, at least about82%, at least about 83%, at least about 84%, at least about 85%, atleast about 86%, at least about 87%, at least about 88%, at least about89%, at least about 90%, at least about 91%, at least about 92%, atleast about 93%, at least about 93%, at least about 94%, at least about95%, at least about 96%, at least about 97%, at least about 98%, atleast about 99%, e.g., 100% identical to the polynucleotide selectedfrom the group consisting of SEQ ID NOs: 619, 617, 606, 615, 629, 1-36,40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616,618, 620-628, 630-638, 642, 645, 650, 651, 670, and 671.

According to some embodiments of the invention the exogenouspolynucleotide is set forth by SEQ ID NO:619, 617, 606, 615, 629, 1-36,40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616,618, 620-628, 630-638, 642, 645, 650, 651, 670, or 671.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing abiotic stress tolerance, yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of a plant, comprising expressing withinthe plant an exogenous polynucleotide comprising the nucleic acidsequence selected from the group consisting of SEQ ID NOs:619, 617, 606,615, 629, 1-49, 51-59, 113-343, 345-351, 353-358, 605, 607-614, 616,618, 620-628, 630-638, 641, 642, 644, 644-646, 648-651, 670, and 671,thereby increasing the abiotic stress tolerance, yield, biomass, growthrate, vigor, oil content, fiber yield, fiber quality, and/or nitrogenuse efficiency of the plant.

According to some embodiments of the invention the exogenouspolynucleotide is set forth by the nucleic acid sequence selected fromthe group consisting of SEQ ID NOs:619, 617, 606, 615, 629, 1-49, 51-59,113-343, 345-351, 353-358, 605, 607-614, 616, 618, 620-628, 630-638,641, 642, 644, 644-646, 648-651, 670, and 671.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing abiotic stress tolerance, nitrogen useefficiency, fiber yield and/or fiber quality of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence at least about 80%, at least about 81%, at leastabout 82%, at least about 83%, at least about 84%, at least about 85%,at least about 86%, at least about 87%, at least about 88%, at leastabout 89%, at least about 90%, at least about 91%, at least about 92%,at least about 93%, at least about 93%, at least about 94%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, e.g., 100% identical to the polynucleotide selectedfrom the group consisting of SEQ ID NOs:352, 639, 640, and 643, therebyincreasing the abiotic stress tolerance, nitrogen use efficiency, fiberyield and/or fiber quality of the plant.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing abiotic stress tolerance, nitrogen useefficiency, fiber yield and/or fiber quality of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising thenucleic acid sequence selected from the group consisting of SEQ ID NOs:352, 639, 640, and 643, thereby increasing the abiotic stress tolerance,nitrogen use efficiency, fiber yield and/or fiber quality of the plant.

According to some embodiments of the invention the exogenouspolynucleotide is set forth by the nucleic acid sequence selected fromthe group consisting of SEQ ID NOs: 352, 639, 640, and 643.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing nitrogen use efficiency, seed yieldand/or oil content of a plant, comprising expressing within the plant anexogenous polynucleotide comprising a nucleic acid sequence at leastabout 80%, at least about 81%, at least about 82%, at least about 83%,at least about 84%, at least about 85%, at least about 86%, at leastabout 87%, at least about 88%, at least about 89%, at least about 90%,at least about 91%, at least about 92%, at least about 93%, at leastabout 93%, at least about 94%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, at least about 99%, e.g., 100%identical to the polynucleotide selected from the group consisting ofSEQ ID NOs:50, 645, and 647, thereby increasing the nitrogen useefficiency, seed yield and/or oil content of the plant.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing nitrogen use efficiency, seed yieldand/or oil content of a plant, comprising expressing within the plant anexogenous polynucleotide comprising the nucleic acid sequence selectedfrom the group consisting of SEQ ID NOs:50, 645 and 647, therebyincreasing the nitrogen use efficiency, seed yield and/or oil content ofthe plant.

According to some embodiments of the invention the exogenouspolynucleotide is set forth by the nucleic acid sequence selected fromthe group consisting of SEQ ID NOs: 50, 645 and 647.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing seed yield, fiber yield and/or fiberquality of a plant, comprising expressing within the plant an exogenouspolynucleotide comprising a nucleic acid sequence at least about 80%, atleast about 81%, at least about 82%, at least about 83%, at least about84%, at least about 85%, at least about 86%, at least about 87%, atleast about 88%, at least about 89%, at least about 90%, at least about91%, at least about 92%, at least about 93%, at least about 93%, atleast about 94%, at least about 95%, at least about 96%, at least about97%, at least about 98%, at least about 99%, e.g., 100% identical to thepolynucleotide set forth by SEQ ID NO:344, thereby increasing the seedyield, fiber yield and/or fiber quality of the plant.

According to an aspect of some embodiments of the invention, there isprovided a method of increasing seed yield, fiber yield and/or fiberquality of a plant, comprising expressing within the plant an exogenouspolynucleotide comprising the nucleic acid sequence set forth in SEQ IDNO:344, thereby increasing the seed yield, fiber yield and/or fiberquality of the plant.

According to some embodiments of the invention the exogenouspolynucleotide is set forth by the nucleic acid sequence set forth inSEQ ID NO:344.

As used herein the term “polynucleotide” refers to a single or doublestranded nucleic acid sequence which is isolated and provided in theform of an RNA sequence, a complementary polynucleotide sequence (cDNA),a genomic polynucleotide sequence and/or a composite polynucleotidesequences (e.g., a combination of the above).

The term “isolated” refers to at least partially separated from thenatural environment e.g., from a plant cell.

As used herein the phrase “complementary polynucleotide sequence” refersto a sequence, which results from reverse transcription of messenger RNAusing a reverse transcriptase or any other RNA dependent DNA polymerase.Such a sequence can be subsequently amplified in vivo or in vitro usinga DNA dependent DNA polymerase.

As used herein the phrase “genomic polynucleotide sequence” refers to asequence derived (isolated) from a chromosome and thus it represents acontiguous portion of a chromosome.

As used herein the phrase “composite polynucleotide sequence” refers toa sequence, which is at least partially complementary and at leastpartially genomic. A composite sequence can include some exonalsequences required to encode the polypeptide of the present invention,as well as some intronic sequences interposing therebetween. Theintronic sequences can be of any source, including of other genes, andtypically will include conserved splicing signal sequences. Suchintronic sequences may further include cis acting expression regulatoryelements.

According to some embodiments of the invention, the exogenouspolynucleotide comprising a nucleic acid sequence encoding a polypeptidehaving an amino acid sequence at least about 80%, at least about 81%, atleast about 82%, at least about 83%, at least about 84%, at least about85%, at least about 86%, at least about 87%, at least about 88%, atleast about 89%, at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or more say 100% homologous to the amino acid sequence selectedfrom the group consisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72,74, 76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, and672.

Homology (e.g., percent homology) can be determined using any homologycomparison software, including for example, the BlastP or TBLASTNsoftware of the National Center of Biotechnology Information (NCBI) suchas by using default parameters, when starting from a polypeptidesequence; or the tBLASTX algorithm (available via the NCBI) such as byusing default parameters, which compares the six-frame conceptualtranslation products of a nucleotide query sequence (both strands)against a protein sequence database.

Homologous sequences include both orthologous and paralogous sequences.The term “paralogous” relates to gene-duplications within the genome ofa species leading to paralogous genes. The term “orthologous” relates tohomologous genes in different organisms due to ancestral relationship.

One option to identify orthologues in monocot plant species is byperforming a reciprocal blast search. This may be done by a first blastinvolving blasting the sequence-of-interest against any sequencedatabase, such as the publicly available NCBI database which may befound at: Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot)nlm (dot) nih (dot) gov. If orthologues in rice were sought, thesequence-of-interest would be blasted against, for example, the 28,469full-length cDNA clones from Oryza sativa Nipponbare available at NCBI.The blast results may be filtered. The full-length sequences of eitherthe filtered results or the non-filtered results are then blasted back(second blast) against the sequences of the organism from which thesequence-of-interest is derived. The results of the first and secondblasts are then compared. An orthologue is identified when the sequenceresulting in the highest score (best hit) in the first blast identifiesin the second blast the query sequence (the originalsequence-of-interest) as the best hit. Using the same rational aparalogue (homolog to a gene in the same organism) is found. In case oflarge sequence families, the ClustalW program may be used [HypertextTransfer Protocol://World Wide Web (dot) ebi (dot) ac (dot)uk/Tools/clustalw2/index (dot) html], followed by a neighbor-joiningtree (Hypertext Transfer Protocol://en (dot) wikipedia (dot)org/wiki/Neighbor-joining) which helps visualizing the clustering.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO: 75, 73, 652, 71, 86, 60-70, 72, 74,76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, or 672.

According to an aspect of some embodiments of the invention, the methodof increasing abiotic stress tolerance, yield, biomass, growth rate,vigor, oil content, fiber yield, fiber quality, and/or nitrogen useefficiency of a plant, is effected by expressing within the plant anexogenous polynucleotide comprising a nucleic acid sequence encoding apolypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72, 74, 76-85,87-98, 100-109, 111, 112, 359-589, 591-597, 600-604, 653-662, 664,666-669, and 672, thereby increasing the abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of the plant.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO: 75, 73, 652, 71, 86, 60-70, 72, 74,76-85, 87-98, 100-109, 111, 112, 359-589, 591-597, 600-604, 653-662,664, 666-669, or 672.

According to an aspect of some embodiments of the invention, the methodof increasing abiotic stress tolerance, nitrogen use efficiency, fiberyield and/or fiber quality of a plant, is effected by expressing withinthe plant an exogenous polynucleotide comprising a nucleic acid sequenceencoding a polypeptide having an amino acid sequence least about 80%, atleast about 81%, at least about 82%, at least about 83%, at least about84%, at least about 85%, at least about 86%, at least about 87%, atleast about 88%, at least about 89%, at least about 90%, at least about91%, at least about 92%, at least about 93%, at least about 94%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or more say 100% homologous to the amino acidsequence selected from the group consisting of SEQ ID NOs:99 and 598,thereby increasing the abiotic stress tolerance, nitrogen useefficiency, fiber yield and/or fiber quality of the plant.

According to an aspect of some embodiments of the invention, the methodof increasing abiotic stress tolerance, nitrogen use efficiency, fiberyield and/or fiber quality of a plant is effected by expressing withinthe plant an exogenous polynucleotide comprising a nucleic acid sequenceencoding a polypeptide comprising the amino acid sequence selected fromthe group consisting of SEQ ID NOs: 99 and 598, thereby increasing theabiotic stress tolerance, nitrogen use efficiency, fiber yield and/orfiber quality of the plant.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO: 99 or 598.

According to an aspect of some embodiments of the invention, the methodof increasing nitrogen use efficiency, seed yield and/or oil content ofa plant is effected expressing within the plant an exogenouspolynucleotide comprising a nucleic acid sequence encoding a polypeptidehaving an amino acid sequence least about 80%, at least about 81%, atleast about 82%, at least about 83%, at least about 84%, at least about85%, at least about 86%, at least about 87%, at least about 88%, atleast about 89%, at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or more say 100% homologous to the amino acid sequence selectedfrom the group consisting of SEQ ID NOs:599 and 663, thereby increasingthe nitrogen use efficiency, seed yield and/or oil content of the plant.

According to an aspect of some embodiments of the invention, the methodof increasing nitrogen use efficiency, seed yield and/or oil content ofa plant is effected by expressing within the plant an exogenouspolynucleotide comprising a nucleic acid sequence encoding a polypeptidecomprising the amino acid sequence selected from the group consisting ofSEQ ID NOs: 599 and 663, thereby increasing the nitrogen use efficiency,seed yield and/or oil content of the plant.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO: 599 or 663.

According to an aspect of some embodiments of the invention, the methodof increasing nitrogen use efficiency, abiotic stress tolerance, seedyield and/or oil content of a plant is effected by expressing within theplant an exogenous polynucleotide comprising a nucleic acid sequenceencoding a polypeptide having an amino acid sequence least about 80%, atleast about 81%, at least about 82%, at least about 83%, at least about84%, at least about 85%, at least about 86%, at least about 87%, atleast about 88%, at least about 89%, at least about 90%, at least about91%, at least about 92%, at least about 93%, at least about 94%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or more say 100% homologous to the amino acidsequence selected from the group consisting of SEQ ID NOs:110 and 665,thereby increasing the nitrogen use efficiency, abiotic stresstolerance, seed yield and/or oil content of the plant.

According to an aspect of some embodiments of the invention, the methodof increasing nitrogen use efficiency, abiotic stress tolerance, seedyield and/or oil content of a plant is effected by expressing within theplant an exogenous polynucleotide comprising a nucleic acid sequenceencoding a polypeptide comprising the amino acid sequence selected fromthe group consisting of SEQ ID NOs:110 and 665, thereby increasing thenitrogen use efficiency, abiotic stress tolerance, seed yield and/or oilcontent of the plant.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO: 110 or 665.

According to an aspect of some embodiments of the invention, the methodof increasing seed yield, fiber yield and/or fiber quality of a plant iseffected by expressing within the plant an exogenous polynucleotidecomprising a nucleic acid sequence encoding a polypeptide having anamino acid sequence least about 80%, at least about 81%, at least about82%, at least about 83%, at least about 84%, at least about 85%, atleast about 86%, at least about 87%, at least about 88%, at least about89%, at least about 90%, at least about 91%, at least about 92%, atleast about 93%, at least about 94%, at least about 95%, at least about96%, at least about 97%, at least about 98%, at least about 99%, or moresay 100% homologous to the amino acid sequence set forth by SEQ IDNO:590, thereby increasing the seed yield, fiber yield and/or fiberquality of the plant.

According to an aspect of some embodiments of the invention, the methodof increasing seed yield, fiber yield and/or fiber quality of a plant iseffected by expressing within the plant an exogenous polynucleotidecomprising a nucleic acid sequence encoding a polypeptide comprising theamino acid sequence set forth by SEQ ID NO:590, thereby increasing theseed yield, fiber yield and/or fiber quality of the plant.

According to some embodiments of the invention, the exogenouspolynucleotide encodes a polypeptide consisting of the amino acidsequence set forth by SEQ ID NO:590.

Nucleic acid sequences encoding the polypeptides of the presentinvention may be optimized for expression. Non-limiting examples ofoptimized nucleic acid sequences are provided in SEQ ID NOs: 670(BDL103_long), 639 (BDL11) and 643 (BDL17) which encode optimizedpolypeptide comprising the amino acid sequences set forth by SEQ ID NOs:96, 661 and 101, respectively. Examples of such sequence modificationsinclude, but are not limited to, an altered G/C content to more closelyapproach that typically found in the plant species of interest, and theremoval of codons atypically found in the plant species commonlyreferred to as codon optimization.

The phrase “codon optimization” refers to the selection of appropriateDNA nucleotides for use within a structural gene or fragment thereofthat approaches codon usage within the plant of interest. Therefore, anoptimized gene or nucleic acid sequence refers to a gene in which thenucleotide sequence of a native or naturally occurring gene has beenmodified in order to utilize statistically-preferred orstatistically-favored codons within the plant. The nucleotide sequencetypically is examined at the DNA level and the coding region optimizedfor expression in the plant species determined using any suitableprocedure, for example as described in Sardana et al. (1996, Plant CellReports 15:677-681). In this method, the standard deviation of codonusage, a measure of codon usage bias, may be calculated by first findingthe squared proportional deviation of usage of each codon of the nativegene relative to that of highly expressed plant genes, followed by acalculation of the average squared deviation. The formula used is: 1SDCU=n=1 N [(Xn−Yn)/Yn]2/N, where Xn refers to the frequency of usage ofcodon n in highly expressed plant genes, where Yn to the frequency ofusage of codon n in the gene of interest and N refers to the totalnumber of codons in the gene of interest. A Table of codon usage fromhighly expressed genes of dicotyledonous plants is compiled using thedata of Murray et al. (1989, Nuc Acids Res. 17:477-498).

One method of optimizing the nucleic acid sequence in accordance withthe preferred codon usage for a particular plant cell type is based onthe direct use, without performing any extra statistical calculations,of codon optimization Tables such as those provided on-line at the CodonUsage Database through the NIAS (National Institute of AgrobiologicalSciences) DNA bank in Japan (Hypertext Transfer Protocol://World WideWeb (dot) kazusa (dot) or (dot) jp/codon/). The Codon Usage Databasecontains codon usage tables for a number of different species, with eachcodon usage Table having been statistically determined based on the datapresent in Genbank.

By using the above Tables to determine the most preferred or mostfavored codons for each amino acid in a particular species (for example,rice), a naturally-occurring nucleotide sequence encoding a protein ofinterest can be codon optimized for that particular plant species. Thisis effected by replacing codons that may have a low statisticalincidence in the particular species genome with corresponding codons, inregard to an amino acid, that are statistically more favored. However,one or more less-favored codons may be selected to delete existingrestriction sites, to create new ones at potentially useful junctions(5′ and 3′ ends to add signal peptide or termination cassettes, internalsites that might be used to cut and splice segments together to producea correct full-length sequence), or to eliminate nucleotide sequencesthat may negatively effect mRNA stability or expression.

The naturally-occurring encoding nucleotide sequence may already, inadvance of any modification, contain a number of codons that correspondto a statistically-favored codon in a particular plant species.Therefore, codon optimization of the native nucleotide sequence maycomprise determining which codons, within the native nucleotidesequence, are not statistically-favored with regards to a particularplant, and modifying these codons in accordance with a codon usage tableof the particular plant to produce a codon optimized derivative. Amodified nucleotide sequence may be fully or partially optimized forplant codon usage provided that the protein encoded by the modifiednucleotide sequence is produced at a level higher than the proteinencoded by the corresponding naturally occurring or native gene.Construction of synthetic genes by altering the codon usage is describedin for example PCT Patent Application 93/07278.

Thus, the invention encompasses nucleic acid sequences describedhereinabove; fragments thereof, sequences hybridizable therewith,sequences homologous thereto, sequences encoding similar polypeptideswith different codon usage, altered sequences characterized bymutations, such as deletion, insertion or substitution of one or morenucleotides, either naturally occurring or man induced, either randomlyor in a targeted fashion.

The invention provides an isolated polynucleotide comprising a nucleicacid sequence at least about 80%, at least about 81%, at least about82%, at least about 83%, at least about 84%, at least about 85%, atleast about 86%, at least about 87%, at least about 88%, at least about89%, at least about 90%, at least about 91%, at least about 92%, atleast about 93%, at least about 93%, at least about 94%, at least about95%, at least about 96%, at least about 97%, at least about 98%, atleast about 99%, e.g., 100% identical to the polynucleotide selectedfrom the group consisting of SEQ ID NOs: 619, 617, 606, 615, 629, 1-36,40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616,618, 620-628, 630-638, 642, 645, 650-651, 670, and 671.

According to some embodiments of the invention the nucleic acid sequenceis capable of increasing abiotic stress tolerance, yield, biomass,growth rate, vigor, oil content, fiber yield, fiber quality, and/ornitrogen use efficiency of a plant.

According to some embodiments of the invention the isolatedpolynucleotide comprising the nucleic acid sequence selected from thegroup consisting of SEQ ID NOs: 619, 617, 606, 615, 629, 1-36, 40, 41,43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616, 618,620-628, 630-638, 642, 645, 650-651, 670, and 671.

According to some embodiments of the invention the isolatedpolynucleotide consists of the nucleic acid sequence selected from thegroup consisting of SEQ ID NOs:619, 617, 606, 615, 629, 1-36, 40, 41,43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616, 618,620-628, 630-638, 642, 645, 650, 651, 670, and 671.

According to some embodiments of the invention the isolatedpolynucleotide is set forth by SEQ ID NO: 619, 617, 606, 615, 629, 1-36,40, 41, 43-45, 49, 52-56, 58, 113-343, 351, 354-358, 605, 607-614, 616,618, 620-628, 630-638, 642, 645, 650-651, 670, and 671.

According to an aspect of some embodiments of the invention, there isprovided an isolated polynucleotide comprising the nucleic acid sequenceselected from the group consisting of SEQ ID NOs:619, 617, 606, 615,629, 1-49, 51-59, 113-343, 345-351, 353-358, 605, 607-614, 616, 618,620-628, 630-638, 641, 642, 644, 644-646, 648-651, 670, and 671.

The invention provides an isolated polynucleotide comprising a nucleicacid sequence encoding a polypeptide which comprises an amino acidsequence at least about 80%, at least about 81%, at least about 82%, atleast about 83%, at least about 84%, at least about 85%, at least about86%, at least about 87%, at least about 88%, at least about 89%, atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or more say 100% homologous to the amino acid sequence selectedfrom the group consisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72,74, 76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, and672.

According to some embodiments of the invention the amino acid sequenceis capable of increasing abiotic stress tolerance, yield, biomass,growth rate, vigor, oil content, fiber yield, fiber quality, and/ornitrogen use efficiency of a plant.

The invention provides an isolated polynucleotide comprising a nucleicacid sequence encoding a polypeptide which comprises the amino acidsequence selected from the group consisting of SEQ ID NOs: 75, 73, 652,71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589,591-597, 600-604, 653-662, 664, 666-669, and 672.

The invention provides an isolated polypeptide comprising an amino acidsequence at least about 80%, at least about 81%, at least about 82%, atleast about 83%, at least about 84%, at least about 85%, at least about86%, at least about 87%, at least about 88%, at least about 89%, atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or more say 100% homologous to an amino acid sequence selected fromthe group consisting of SEQ ID NOs:75, 73, 652, 71, 86, 60-70, 72, 74,76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, and672.

According to some embodiments of the invention the isolated polypeptideis capable of increasing abiotic stress tolerance, yield, biomass,growth rate, vigor, oil content, fiber yield, fiber quality, and/ornitrogen use efficiency of a plant.

According to some embodiments of the invention, the polypeptidecomprising an amino acid sequence selected from the group consisting ofSEQ ID NOs: 75, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109,111, 112, 359-589, 591-597, 600-604, 653-662, 664, 666-669, and 672.

According to some embodiments of the invention, the polypeptide is setforth by SEQ ID NO: 75, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-98,100-109, 111, 112, 359-589, 591-597, 600-604, 653-662, 664, 666-669, or672.

The invention also encompasses fragments of the above describedpolypeptides and polypeptides having mutations, such as deletions,insertions or substitutions of one or more amino acids, either naturallyoccurring or man induced, either randomly or in a targeted fashion.

The term “plant” as used herein encompasses whole plants, ancestors andprogeny of the plants and plant parts, including seeds, shoots, stems,roots (including tubers), and plant cells, tissues and organs. The plantmay be in any form including suspension cultures, embryos, meristematicregions, callus tissue, leaves, gametophytes, sporophytes, pollen, andmicrospores. Plants that are particularly useful in the methods of theinvention include all plants which belong to the superfamilyViridiplantae, in particular monocotyledonous and dicotyledonous plantsincluding a fodder or forage legume, ornamental plant, food crop, tree,or shrub selected from the list comprising Acacia spp., Acer spp.,Actinidia spp., Aesculus spp., Agathis australis, Albizia amara,Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Asteliafragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassicaspp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadabafarinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicumspp., Cassia spp., Centroema pubescens, Chacoomeles spp., Cinnamomumcassia, Coffea arabica, Colophospermum mopane, Coronillia varia,Cotoneaster serotina, Crataegus spp., Cucumis spp., Cupressus spp.,Cyathea dealbata, Cyclonia oblonga, Cryptomeria japonica, Cymbopogonspp., Cynthea dealbata, Cyclonia oblonga, Dalbergia monetaria, Davalliadivaricata, Desmodium spp., Dicksonia squarosa, Dibeteropogonamplectens, Dioclea spp, Dolichos spp., Dorycnium rectum, Echinochloapyramidalis, Ehraffia spp., Eleusine coracana, Eragrestis spp.,Erythrina spp., Eucalypfus spp., Euclea schimperi, Eulalia vi/losa,Pagopyrum spp., Feijoa sellowlana, Fragaria spp., Flemingia spp,Freycinetia banksli, Geranium thunbergii, GinAgo biloba, Glycinejavanica, Gliricidia spp, Gossypium hirsutum, Grevillea spp., Guibourtiacoleosperma, Hedysarum spp., Hemaffhia altissima, Heteropogon contoffus,Hordeum vulgare, Hyparrhenia rufa, Hypericum erectum, Hypeffheliadissolute, Indigo incamata, Iris spp., Leptarrhena pyrolifolia,Lespediza spp., Lettuca spp., Leucaena leucocephala, Loudetia simplex,Lotonus bainesli, Lotus spp., Macrotyloma axillare, Malus spp., Manihotesculenta, Medicago saliva, Metasequoia glyptostroboides, Musasapientum, Nicotianum spp., Onobrychis spp., Ornithopus spp., Oryzaspp., Peltophorum africanum, Pennisetum spp., Persea gratissima, Petuniaspp., Phaseolus spp., Phoenix canariensis, Phormium cookianum, Photiniaspp., Picea glauca, Pinus spp., Pisum sativam, Podocarpus totara,Pogonarthria fleckii, Pogonaffhria squarrosa, Populus spp., Prosopiscineraria, Pseudotsuga menziesii, Pterolobium stellatum, Pyrus communis,Quercus spp., Rhaphiolepsis umbellata, Rhopalostylis sapida, Rhusnatalensis, Ribes grossularia, Ribes spp., Robinia pseudoacacia, Rosaspp., Rubus spp., Salix spp., Schyzachyrium sanguineum, Sciadopitysvefficillata, Sequoia sempervirens, Sequoiadendron giganteum, Sorghumbicolor, Spinacia spp., Sporobolus fimbriatus, Stiburus alopecuroides,Stylosanthos humilis, Tadehagi spp, Taxodium distichum, Themedatriandra, Trifolium spp., Triticum spp., Tsuga heterophylla, Vacciniumspp., Vicia spp., Vitis vinifera, Watsonia pyramidata, Zantedeschiaaethiopica, Zea mays, amaranth, artichoke, asparagus, broccoli, Brusselssprouts, cabbage, canola, carrot, cauliflower, celery, collard greens,flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean,straw, sugar beet, sugar cane, sunflower, tomato, squash tea, maize,wheat, barely, rye, oat, peanut, pea, lentil and alfalfa, cotton,rapeseed, canola, pepper, sunflower, tobacco, eggplant, eucalyptus, atree, an ornamental plant, a perennial grass and a forage crop.Alternatively algae and other non-Viridiplantae can be used for themethods of the present invention.

According to some embodiments of the invention, the plant used by themethod of the invention is a crop plant such as rice, maize, wheat,barley, peanut, potato, sesame, olive tree, palm oil, banana, soybean,sunflower, canola, sugarcane, alfalfa, millet, leguminosae (bean, pea),flax, lupinus, rapeseed, tobacco, poplar and cotton.

According to some embodiments of the invention, there is provided aplant cell exogenously expressing the polynucleotide of some embodimentsof the invention, the nucleic acid construct of some embodiments of theinvention and/or the polypeptide of some embodiments of the invention.

According to some embodiments of the invention, expressing the exogenouspolynucleotide of the invention within the plant is effected bytransforming one or more cells of the plant with the exogenouspolynucleotide, followed by generating a mature plant from thetransformed cells and cultivating the mature plant under conditionssuitable for expressing the exogenous polynucleotide within the matureplant.

According to some embodiments of the invention, the transformation iseffected by introducing to the plant cell a nucleic acid construct whichincludes the exogenous polynucleotide of some embodiments of theinvention and at least one promoter for directing transcription of theexogenous polynucleotide in a host cell (a plant cell). Further detailsof suitable transformation approaches are provided hereinbelow.

According to some embodiments of the invention, there is provided anucleic acid construct comprising the isolated polynucleotide of theinvention, and a promoter for directing transcription of the nucleicacid sequence of the isolated polynucleotide in a host cell.

According to some embodiments of the invention, the isolatedpolynucleotide is operably linked to the promoter sequence.

A coding nucleic acid sequence is “operably linked” to a regulatorysequence (e.g., promoter) if the regulatory sequence is capable ofexerting a regulatory effect on the coding sequence linked thereto.

As used herein, the term “promoter” refers to a region of DNA which liesupstream of the transcriptional initiation site of a gene to which RNApolymerase binds to initiate transcription of RNA. The promoter controlswhere (e.g., which portion of a plant) and/or when (e.g., at which stageor condition in the lifetime of an organism) the gene is expressed.

Any suitable promoter sequence can be used by the nucleic acid constructof the present invention. Preferably the promoter is a constitutivepromoter, a tissue-specific, or an abiotic stress-inducible promoter.

Suitable constitutive promoters include, for example, CaMV 35S promoter(SEQ ID NO:675; Odell et al., Nature 313:810-812, 1985); ArabidopsisAt6669 promoter (SEQ ID NO:674; see PCT Publication No. WO04081173A2);maize Ubi 1 (Christensen et al., Plant Sol. Biol. 18:675-689, 1992);rice actin (McElroy et al., Plant Cell 2:163-171, 1990); pEMU (Last etal., Theor. Appl. Genet. 81:581-588, 1991); CaMV 19S (Nilsson et al.,Physiol. Plant 100:456-462, 1997); GOS2 (de Pater et al, Plant JNovember;2(6):837-44, 1992); ubiquitin (Christensen et al, Plant Mol.Biol. 18: 675-689, 1992); Rice cyclophilin (Bucholz et al, Plant Mol.Biol. 25(5):837-43, 1994); Maize H3 histone (Lepetit et al, Mol. Gen.Genet. 231: 276-285, 1992); Actin 2 (An et al, Plant J. 10(1); 107-121,1996) and Synthetic Super MAS (Ni et al., The Plant Journal 7: 661-76,1995). Other constitutive promoters include those in U.S. Pat. Nos.5,659,026, 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785;5,399,680; 5,268,463; and U.S. Pat. No. 5,608,142.

Suitable tissue-specific promoters include, but not limited to,leaf-specific promoters [such as described, for example, by Yamamoto etal., Plant J. 12:255-265, 1997; Kwon et al., Plant Physiol. 105:357-67,1994; Yamamoto et al., Plant Cell Physiol. 35:773-778, 1994; Gotor etal., Plant J. 3:509-18, 1993; Orozco et al., Plant Mol. Biol.23:1129-1138, 1993; and Matsuoka et al., Proc. Natl. Acad. Sci. USA90:9586-9590, 1993], seed-preferred promoters [e.g., from seed specificgenes (Simon, et al., Plant Mol. Biol. 5. 191, 1985; Scofield, et al.,J. Biol. Chem. 262: 12202, 1987; Baszczynski, et al., Plant Mol. Biol.14: 633, 1990), Brazil Nut albumin (Pearson' et al., Plant Mol. Biol.18: 235-245, 1992), legumin (Ellis, et al. Plant Mol. Biol. 10: 203-214,1988), Glutelin (rice) (Takaiwa, et al., Mol. Gen. Genet. 208: 15-22,1986; Takaiwa, et al., FEBS Letts. 221: 43-47, 1987), Zein (Matzke et alPlant Mol Biol, 143).323-32 1990), napA (Stalberg, et al, Planta 199:515-519, 1996), Wheat SPA (Albanietal, Plant Cell, 9: 171-184, 1997),sunflower oleosin (Cummins, et al., Plant Mol. Biol. 19: 873-876,1992)], endosperm specific promoters [e.g., wheat LMW and HMW,glutenin-1 (Mol Gen Genet. 216:81-90, 1989; NAR 17:461-2), wheat a, band g gliadins (EMBO3:1409-15, 1984), Barley ltrl promoter, barley B1,C, D hordein (Theor Appl Gen 98:1253-62, 1999; Plant J 4:343-55, 1993;Mol Gen Genet. 250:750-60, 1996), Barley DOF (Mena et al, The PlantJournal, 116(1): 53-62, 1998), Biz2 (EP99106056.7), Synthetic promoter(Vicente-Carbajosa et al., Plant J. 13: 629-640, 1998), rice prolaminNRP33, rice-globulin Glb-1 (Wu et al, Plant Cell Physiology 39(8)885-889, 1998), rice alpha-globulin REB/OHP-1 (Nakase et al. Plant Mol.Biol. 33: 513-S22, 1997), rice ADP-glucose PP (Trans Res 6:157-68,1997), maize ESR gene family (Plant J 12:235-46, 1997), sorgumgamma-kafirin (PMB 32:1029-35, 1996)], embryo specific promoters [e.g.,rice OSH1 (Sato et al, Proc. Natl. Acad. Sci. USA, 93: 8117-8122), KNOX(Postma-Haarsma of al, Plant Mol. Biol. 39:257-71, 1999), rice oleosin(Wu et at, J. Biochem., 123:386, 1998)], and flower-specific promoters[e.g., AtPRP4, chalene synthase (chsA) (Van der Meer, et al., Plant Mol.Biol. 15, 95-109, 1990), LAT52 (Twell et al Mol. Gen. Genet.217:240-245; 1989), apetala-3].

Suitable abiotic stress-inducible promoters include, but not limited to,salt-inducible promoters such as RD29A (Yamaguchi-Shinozalei et al.,Mol. Gen. Genet. 236:331-340, 1993); drought-inducible promoters such asmaize rab 17 gene promoter (Pla et. al., Plant Mol. Biol. 21:259-266,1993), maize rab28 gene promoter (Busk et. al., Plant J. 11:1285-1295,1997) and maize Ivr2 gene promoter (Pelleschi et. al., Plant Mol. Biol.39:373-380, 1999); heat-inducible promoters such as heat tomatohsp80-promoter from tomato (U.S. Pat. No. 5,187,267).

The nucleic acid construct of some embodiments of the invention canfurther include an appropriate selectable marker and/or an origin ofreplication. According to some embodiments of the invention, the nucleicacid construct utilized is a shuttle vector, which can propagate both inE. coli (wherein the construct comprises an appropriate selectablemarker and origin of replication) and be compatible with propagation incells. The construct according to the present invention can be, forexample, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus oran artificial chromosome.

The nucleic acid construct of some embodiments of the invention can beutilized to stably or transiently transform plant cells. In stabletransformation, the exogenous polynucleotide is integrated into theplant genome and as such it represents a stable and inherited trait. Intransient transformation, the exogenous polynucleotide is expressed bythe cell transformed but it is not integrated into the genome and assuch it represents a transient trait.

There are various methods of introducing foreign genes into bothmonocotyledonous and dicotyledonous plants (Potrykus, I., Annu. Rev.Plant. Physiol., Plant. Mol. Biol. (1991) 42:205-225; Shimamoto et al.,Nature (1989) 338:274-276).

The principle methods of causing stable integration of exogenous DNAinto plant genomic DNA include two main approaches:

(i) Agrobacterium-mediated gene transfer: Klee et al. (1987) Annu. Rev.Plant Physiol. 38:467-486; Klee and Rogers in Cell Culture and SomaticCell Genetics of Plants, Vol. 6, Molecular Biology of Plant NuclearGenes, eds. Schell, J., and Vasil, L. K., Academic Publishers, SanDiego, Calif. (1989) p. 2-25; Gatenby, in Plant Biotechnology, eds.Kung, S, and Arntzen, C. J., Butterworth Publishers, Boston, Mass.(1989) p. 93-112.

(ii) Direct DNA uptake: Paszkowski et al., in Cell Culture and SomaticCell Genetics of Plants, Vol. 6, Molecular Biology of Plant NuclearGenes eds. Schell, J., and Vasil, L. K., Academic Publishers, San Diego,Calif. (1989) p. 52-68; including methods for direct uptake of DNA intoprotoplasts, Toriyama, K. et al. (1988) Bio/Technology 6:1072-1074. DNAuptake induced by brief electric shock of plant cells: Zhang et al.Plant Cell Rep. (1988) 7:379-384. Fromm et al. Nature (1986)319:791-793. DNA injection into plant cells or tissues by particlebombardment, Klein et al. Bio/Technology (1988) 6:559-563; McCabe et al.Bio/Technology (1988) 6:923-926; Sanford, Physiol. Plant. (1990)79:206-209; by the use of micropipette systems: Neuhaus et al., Theor.Appl. Genet. (1987) 75:30-36; Neuhaus and Spangenberg, Physiol. Plant.(1990) 79:213-217; glass fibers or silicon carbide whiskertransformation of cell cultures, embryos or callus tissue, U.S. Pat. No.5,464,765 or by the direct incubation of DNA with germinating pollen,DeWet et al. in Experimental Manipulation of Ovule Tissue, eds. Chapman,G. P. and Mantell, S. H. and Daniels, W. Longman, London, (1985) p.197-209; and Ohta, Proc. Natl. Acad. Sci. USA (1986) 83:715-719.

The Agrobacterium system includes the use of plasmid vectors thatcontain defined DNA segments that integrate into the plant genomic DNA.Methods of inoculation of the plant tissue vary depending upon the plantspecies and the Agrobacterium delivery system. A widely used approach isthe leaf disc procedure which can be performed with any tissue explantthat provides a good source for initiation of whole plantdifferentiation. See, e.g., Horsch et al. in Plant Molecular BiologyManual A5, Kluwer Academic Publishers, Dordrecht (1988) p. 1-9. Asupplementary approach employs the Agrobacterium delivery system incombination with vacuum infiltration. The Agrobacterium system isespecially viable in the creation of transgenic dicotyledonous plants.

There are various methods of direct DNA transfer into plant cells. Inelectroporation, the protoplasts are briefly exposed to a strongelectric field. In microinjection, the DNA is mechanically injecteddirectly into the cells using very small micropipettes. In microparticlebombardment, the DNA is adsorbed on microprojectiles such as magnesiumsulfate crystals or tungsten particles, and the microprojectiles arephysically accelerated into cells or plant tissues.

Following stable transformation plant propagation is exercised. The mostcommon method of plant propagation is by seed. Regeneration by seedpropagation, however, has the deficiency that due to heterozygositythere is a lack of uniformity in the crop, since seeds are produced byplants according to the genetic variances governed by Mendelian rules.Basically, each seed is genetically different and each will grow withits own specific traits. Therefore, it is preferred that the transformedplant be produced such that the regenerated plant has the identicaltraits and characteristics of the parent transgenic plant. Therefore, itis preferred that the transformed plant be regenerated bymicropropagation which provides a rapid, consistent reproduction of thetransformed plants.

Micropropagation is a process of growing new generation plants from asingle piece of tissue that has been excised from a selected parentplant or cultivar. This process permits the mass reproduction of plantshaving the preferred tissue expressing the fusion protein. The newgeneration plants which are produced are genetically identical to, andhave all of the characteristics of, the original plant. Micropropagationallows mass production of quality plant material in a short period oftime and offers a rapid multiplication of selected cultivars in thepreservation of the characteristics of the original transgenic ortransformed plant. The advantages of cloning plants are the speed ofplant multiplication and the quality and uniformity of plants produced.

Micropropagation is a multi-stage procedure that requires alteration ofculture medium or growth conditions between stages. Thus, themicropropagation process involves four basic stages: Stage one, initialtissue culturing; stage two, tissue culture multiplication; stage three,differentiation and plant formation; and stage four, greenhouseculturing and hardening. During stage one, initial tissue culturing, thetissue culture is established and certified contaminant-free. Duringstage two, the initial tissue culture is multiplied until a sufficientnumber of tissue samples are produced to meet production goals. Duringstage three, the tissue samples grown in stage two are divided and growninto individual plantlets. At stage four, the transformed plantlets aretransferred to a greenhouse for hardening where the plants' tolerance tolight is gradually increased so that it can be grown in the naturalenvironment.

According to some embodiments of the invention, the transgenic plantsare generated by transient transformation of leaf cells, meristematiccells or the whole plant.

Transient transformation can be effected by any of the direct DNAtransfer methods described above or by viral infection using modifiedplant viruses.

Viruses that have been shown to be useful for the transformation ofplant hosts include CaMV, Tobacco mosaic virus (TMV), brome mosaic virus(BMV) and Bean Common Mosaic Virus (BV or BCMV). Transformation ofplants using plant viruses is described in U.S. Pat. No. 4,855,237 (beangolden mosaic virus; BGV), EP-A 67,553 (TMV), Japanese PublishedApplication No. 63-14693 (TMV), EPA 194,809 (BV), EPA 278,667 (BV); andGluzman, Y. et al., Communications in Molecular Biology: Viral Vectors,Cold Spring Harbor Laboratory, New York, pp. 172-189 (1988). Pseudovirusparticles for use in expressing foreign DNA in many hosts, includingplants are described in WO 87/06261.

According to some embodiments of the invention, the virus used fortransient transformations is avirulent and thus is incapable of causingsevere symptoms such as reduced growth rate, mosaic, ring spots, leafroll, yellowing, streaking, pox formation, tumor formation and pitting.A suitable avirulent virus may be a naturally occurring avirulent virusor an artificially attenuated virus. Virus attenuation may be effectedby using methods well known in the art including, but not limited to,sub-lethal heating, chemical treatment or by directed mutagenesistechniques such as described, for example, by Kurihara and Watanabe(Molecular Plant Pathology 4:259-269, 2003), Gal-on et al. (1992),Atreya et al. (1992) and Huet et al. (1994).

Suitable virus strains can be obtained from available sources such as,for example, the American Type culture Collection (ATCC) or by isolationfrom infected plants. Isolation of viruses from infected plant tissuescan be effected by techniques well known in the art such as described,for example by Foster and Tatlor, Eds. “Plant Virology Protocols FromVirus Isolation to Transgenic Resistance (Methods in Molecular Biology(Humana Pr), Vol 81)”, Humana Press, 1998. Briefly, tissues of aninfected plant believed to contain a high concentration of a suitablevirus, preferably young leaves and flower petals, are ground in a buffersolution (e.g., phosphate buffer solution) to produce a virus infectedsap which can be used in subsequent inoculations.

Construction of plant RNA viruses for the introduction and expression ofnon-viral exogenous polynucleotide sequences in plants is demonstratedby the above references as well as by Dawson, W. O. et al., Virology(1989) 172:285-292; Takamatsu et al. EMBO J. (1987) 6:307-311; French etal. Science (1986) 231:1294-1297; Takamatsu et al. FEBS Letters (1990)269:73-76; and U.S. Pat. No. 5,316,931.

When the virus is a DNA virus, suitable modifications can be made to thevirus itself. Alternatively, the virus can first be cloned into abacterial plasmid for ease of constructing the desired viral vector withthe foreign DNA. The virus can then be excised from the plasmid. If thevirus is a DNA virus, a bacterial origin of replication can be attachedto the viral DNA, which is then replicated by the bacteria.Transcription and translation of this DNA will produce the coat proteinwhich will encapsidate the viral DNA. If the virus is an RNA virus, thevirus is generally cloned as a cDNA and inserted into a plasmid. Theplasmid is then used to make all of the constructions. The RNA virus isthen produced by transcribing the viral sequence of the plasmid andtranslation of the viral genes to produce the coat protein(s) whichencapsidate the viral RNA.

In one embodiment, a plant viral polynucleotide is provided in which thenative coat protein coding sequence has been deleted from a viralpolynucleotide, a non-native plant viral coat protein coding sequenceand a non-native promoter, preferably the subgenomic promoter of thenon-native coat protein coding sequence, capable of expression in theplant host, packaging of the recombinant plant viral polynucleotide, andensuring a systemic infection of the host by the recombinant plant viralpolynucleotide, has been inserted. Alternatively, the coat protein genemay be inactivated by insertion of the non-native polynucleotidesequence within it, such that a protein is produced. The recombinantplant viral polynucleotide may contain one or more additional non-nativesubgenomic promoters. Each non-native subgenomic promoter is capable oftranscribing or expressing adjacent genes or polynucleotide sequences inthe plant host and incapable of recombination with each other and withnative subgenomic promoters. Non-native (foreign) polynucleotidesequences may be inserted adjacent the native plant viral subgenomicpromoter or the native and a non-native plant viral subgenomic promotersif more than one polynucleotide sequence is included. The non-nativepolynucleotide sequences are transcribed or expressed in the host plantunder control of the subgenomic promoter to produce the desiredproducts.

In a second embodiment, a recombinant plant viral polynucleotide isprovided as in the first embodiment except that the native coat proteincoding sequence is placed adjacent one of the non-native coat proteinsubgenomic promoters instead of a non-native coat protein codingsequence.

In a third embodiment, a recombinant plant viral polynucleotide isprovided in which the native coat protein gene is adjacent itssubgenomic promoter and one or more non-native subgenomic promoters havebeen inserted into the viral polynucleotide. The inserted non-nativesubgenomic promoters are capable of transcribing or expressing adjacentgenes in a plant host and are incapable of recombination with each otherand with native subgenomic promoters. Non-native polynucleotidesequences may be inserted adjacent the non-native subgenomic plant viralpromoters such that the sequences are transcribed or expressed in thehost plant under control of the subgenomic promoters to produce thedesired product.

In a fourth embodiment, a recombinant plant viral polynucleotide isprovided as in the third embodiment except that the native coat proteincoding sequence is replaced by a non-native coat protein codingsequence.

The viral vectors are encapsidated by the coat proteins encoded by therecombinant plant viral polynucleotide to produce a recombinant plantvirus. The recombinant plant viral polynucleotide or recombinant plantvirus is used to infect appropriate host plants. The recombinant plantviral polynucleotide is capable of replication in the host, systemicspread in the host, and transcription or expression of foreign gene(s)(exogenous polynucleotide) in the host to produce the desired protein.

Techniques for inoculation of viruses to plants may be found in Fosterand Taylor, eds. “Plant Virology Protocols: From Virus Isolation toTransgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol81)”, Humana Press, 1998; Maramorosh and Koprowski, eds. “Methods inVirology” 7 vols, Academic Press, New York 1967-1984; Hill, S. A.“Methods in Plant Virology”, Blackwell, Oxford, 1984; Walkey, D. G. A.“Applied Plant Virology”, Wiley, New York, 1985; and Kado and Agrawa,eds. “Principles and Techniques in Plant Virology”, VanNostrand-Reinhold, New York.

In addition to the above, the polynucleotide of the present inventioncan also be introduced into a chloroplast genome thereby enablingchloroplast expression.

A technique for introducing exogenous polynucleotide sequences to thegenome of the chloroplasts is known. This technique involves thefollowing procedures. First, plant cells are chemically treated so as toreduce the number of chloroplasts per cell to about one. Then, theexogenous polynucleotide is introduced via particle bombardment into thecells with the aim of introducing at least one exogenous polynucleotidemolecule into the chloroplasts. The exogenous polynucleotides selectedsuch that it is integratable into the chloroplast's genome viahomologous recombination which is readily effected by enzymes inherentto the chloroplast. To this end, the exogenous polynucleotide includes,in addition to a gene of interest, at least one polynucleotide stretchwhich is derived from the chloroplast's genome. In addition, theexogenous polynucleotide includes a selectable marker, which serves bysequential selection procedures to ascertain that all or substantiallyall of the copies of the chloroplast genomes following such selectionwill include the exogenous polynucleotide. Further details relating tothis technique are found in U.S. Pat. No. 4,945,050; and U.S. Pat. No.5,693,507 which are incorporated herein by reference. A polypeptide canthus be produced by the protein expression system of the chloroplast andbecome integrated into the chloroplast's inner membrane.

Since processes which increase oil content, yield, growth rate, biomass,vigor and/or abiotic stress tolerance of a plant can involve multiplegenes acting additively or in synergy (see, for example, in Quesda etal., Plant Physiol. 130:951-063, 2002), the present invention alsoenvisages expressing a plurality of exogenous polynucleotides in asingle host plant to thereby achieve superior effect on oil content,yield, growth rate, biomass, vigor and/or abiotic stress tolerance.

Expressing a plurality of exogenous polynucleotides in a single hostplant can be effected by co-introducing multiple nucleic acidconstructs, each including a different exogenous polynucleotide, into asingle plant cell. The transformed cell can than be regenerated into amature plant using the methods described hereinabove.

Alternatively, expressing a plurality of exogenous polynucleotides in asingle host plant can be effected by co-introducing into a singleplant-cell a single nucleic-acid construct including a plurality ofdifferent exogenous polynucleotides. Such a construct can be designedwith a single promoter sequence which can transcribe a polycistronicmessenger RNA including all the different exogenous polynucleotidesequences. To enable co-translation of the different polypeptidesencoded by the polycistronic messenger RNA, the polynucleotide sequencescan be inter-linked via an internal ribosome entry site (IRES) sequencewhich facilitates translation of polynucleotide sequences positioneddownstream of the IRES sequence. In this case, a transcribedpolycistronic RNA molecule encoding the different polypeptides describedabove will be translated from both the capped 5′ end and the twointernal IRES sequences of the polycistronic RNA molecule to therebyproduce in the cell all different polypeptides. Alternatively, theconstruct can include several promoter sequences each linked to adifferent exogenous polynucleotide sequence.

The plant cell transformed with the construct including a plurality ofdifferent exogenous polynucleotides, can be regenerated into a matureplant, using the methods described hereinabove.

Alternatively, expressing a plurality of exogenous polynucleotides in asingle host plant can be effected by introducing different nucleic acidconstructs, including different exogenous polynucleotides, into aplurality of plants. The regenerated transformed plants can then becross-bred and resultant progeny selected for superior abiotic stresstolerance, water use efficiency, fertilizer use efficiency, growth,biomass, yield and/or vigor traits, using conventional plant breedingtechniques.

According to some embodiments of the invention, the method furthercomprising growing the plant expressing the exogenous polynucleotideunder the abiotic stress.

Non-limiting examples of abiotic stress conditions include, salinity,drought, water deprivation, excess of water (e.g., flood, waterlogging),etiolation, low temperature, high temperature, heavy metal toxicity,anaerobiosis, nutrient deficiency, nutrient excess, atmosphericpollution and UV irradiation.

Thus, the invention encompasses plants exogenously expressing thepolynucleotide(s), the nucleic acid constructs and/or polypeptide(s) ofthe invention. Once expressed within the plant cell or the entire plant,the level of the polypeptide encoded by the exogenous polynucleotide canbe determined by methods well known in the art such as, activity assays,Western blots using antibodies capable of specifically binding thepolypeptide, Enzyme-Linked Immuno Sorbent Assay (ELISA),radio-immunoassays (RIA), immunohistochemistry, immunocytochemistry,immunofluorescence and the like.

Methods of determining the level in the plant of the RNA transcribedfrom the exogenous polynucleotide are well known in the art and include,for example, Northern blot analysis, reverse transcription polymerasechain reaction (RT-PCR) analysis (including quantitative,semi-quantitative or real-time RT-PCR) and RNA-in situ hybridization.

The sequence information and annotations uncovered by the presentteachings can be harnessed in favor of classical breeding. Thus,sub-sequence data of those polynucleotides described above, can be usedas markers for marker assisted selection (MAS), in which a marker isused for indirect selection of a genetic determinant or determinants ofa trait of interest (e.g., abiotic stress tolerance, increased yield,biomass, growth rate, vigor, oil content, fiber yield, fiber quality,and/or nitrogen use efficiency of a plant). Nucleic acid data of thepresent teachings (DNA or RNA sequence) may contain or be linked topolymorphic sites or genetic markers on the genome such as restrictionfragment length polymorphism (RFLP), micro-satellites and singlenucleotide polymorphism (SNP), DNA fingerprinting (DFP), amplifiedfragment length polymorphism (AFLP), expression level polymorphism,polymorphism of the encoded polypeptide and any other polymorphism atthe DNA or RNA sequence.

Examples of marker assisted selections include, but are not limited to,selection for a morphological trait (e.g., a gene that affects form,coloration, male sterility or resistance such as the presence or absenceof awn, leaf sheath coloration, height, grain color, aroma of rice);selection for a biochemical trait (e.g., a gene that encodes a proteinthat can be extracted and observed; for example, isozymes and storageproteins); selection for a biological trait (e.g., pathogen races orinsect biotypes based on host pathogen or host parasite interaction canbe used as a marker since the genetic constitution of an organism canaffect its susceptibility to pathogens or parasites).

The polynucleotides and polypeptides described hereinabove can be usedin a wide range of economical plants, in a safe and cost effectivemanner.

Plant lines exogenously expressing the polynucleotide or the polypeptideof the invention are screened to identify those that show the greatestincrease of the desired plant trait.

The effect of the transgene (the exogenous polynucleotide encoding thepolypeptide) on abiotic stress tolerance can be determined using knownmethods such as detailed below and in the Examples section whichfollows.

Abiotic Stress Tolerance—

Transformed (i.e., expressing the transgene) and non-transformed (wildtype) plants are exposed to an abiotic stress condition, such as waterdeprivation, suboptimal temperature (low temperature, high temperature),nutrient deficiency, nutrient excess, a salt stress condition, osmoticstress, heavy metal toxicity, anaerobiosis, atmospheric pollution and UVirradiation.

Salinity Tolerance Assay—

Transgenic plants with tolerance to high salt concentrations areexpected to exhibit better germination, seedling vigor or growth in highsalt. Salt stress can be effected in many ways such as, for example, byirrigating the plants with a hyperosmotic solution, by cultivating theplants hydroponically in a hyperosmotic growth solution (e.g., Hoaglandsolution), or by culturing the plants in a hyperosmotic growth medium[e.g., 50% Murashige-Skoog medium (MS medium)]. Since different plantsvary considerably in their tolerance to salinity, the salt concentrationin the irrigation water, growth solution, or growth medium can beadjusted according to the specific characteristics of the specific plantcultivar or variety, so as to inflict a mild or moderate effect on thephysiology and/or morphology of the plants (for guidelines as toappropriate concentration see, Bernstein and Kafkafi, Root Growth UnderSalinity Stress In: Plant Roots, The Hidden Half 3rd ed. Waisel Y, EshelA and Kafkafi U. (editors) Marcel Dekker Inc., New York, 2002, andreference therein).

For example, a salinity tolerance test can be performed by irrigatingplants at different developmental stages with increasing concentrationsof sodium chloride (for example 50 mM, 100 mM, 200 mM, 400 mM NaCl)applied from the bottom and from above to ensure even dispersal of salt.Following exposure to the stress condition the plants are frequentlymonitored until substantial physiological and/or morphological effectsappear in wild type plants. Thus, the external phenotypic appearance,degree of wilting and overall success to reach maturity and yieldprogeny are compared between control and transgenic plants.

Quantitative parameters of tolerance measured include, but are notlimited to, the average wet and dry weight, growth rate, leaf size, leafcoverage (overall leaf area), the weight of the seeds yielded, theaverage seed size and the number of seeds produced per plant.Transformed plants not exhibiting substantial physiological and/ormorphological effects, or exhibiting higher biomass than wild-typeplants, are identified as abiotic stress tolerant plants.

Osmotic Tolerance Test—

Osmotic stress assays (including sodium chloride and mannitol assays)are conducted to determine if an osmotic stress phenotype was sodiumchloride-specific or if it was a general osmotic stress relatedphenotype. Plants which are tolerant to osmotic stress may have moretolerance to drought and/or freezing. For salt and osmotic stressgermination experiments, the medium is supplemented for example with 50mM, 100 mM, 200 mM NaCl or 100 mM, 200 mM NaCl, 400 mM mannitol.

Drought Tolerance Assay/Osmoticum Assay—

Tolerance to drought is performed to identify the genes conferringbetter plant survival after acute water deprivation. To analyze whetherthe transgenic plants are more tolerant to drought, an osmotic stressproduced by the non-ionic osmolyte sorbitol in the medium can beperformed. Control and transgenic plants are germinated and grown inplant-agar plates for 4 days, after which they are transferred to platescontaining 500 mM sorbitol. The treatment causes growth retardation,then both control and transgenic plants are compared, by measuring plantweight (wet and dry), yield, and by growth rates measured as time toflowering.

Conversely, soil-based drought screens are performed with plantsoverexpressing the polynucleotides detailed above. Seeds from controlArabidopsis plants, or other transgenic plants overexpressing thepolypeptide of the invention are germinated and transferred to pots.Drought stress is obtained after irrigation is ceased accompanied byplacing the pots on absorbent paper to enhance the soil-drying rate.Transgenic and control plants are compared to each other when themajority of the control plants develop severe wilting. Plants arere-watered after obtaining a significant fraction of the control plantsdisplaying a severe wilting. Plants are ranked comparing to controls foreach of two criteria: tolerance to the drought conditions and recovery(survival) following re-watering.

Cold Stress Tolerance—

To analyze cold stress, mature (25 day old) plants are transferred to 4°C. chambers for 1 or 2 weeks, with constitutive light. Later on plantsare moved back to greenhouse. Two weeks later damages from chillingperiod, resulting in growth retardation and other phenotypes, arecompared between both control and transgenic plants, by measuring plantweight (wet and dry), and by comparing growth rates measured as time toflowering, plant size, yield, and the like.

Heat Stress Tolerance—

Heat stress tolerance is achieved by exposing the plants to temperaturesabove 34° C. for a certain period. Plant tolerance is examined aftertransferring the plants back to 22° C. for recovery and evaluation after5 days relative to internal controls (non-transgenic plants) or plantsnot exposed to neither cold or heat stress.

Water Use Efficiency—

can be determined as the biomass produced per unit transpiration. Toanalyze WUE, leaf relative water content can be measured in control andtransgenic plants. Fresh weight (FW) is immediately recorded; thenleaves are soaked for 8 hours in distilled water at room temperature inthe dark, and the turgid weight (TW) is recorded. Total dry weight (DW)is recorded after drying the leaves at 60° C. to a constant weight.Relative water content (RWC) is calculated according to the followingFormula I:

RWC=[(FW−DW)/(TW−DW)]×100  Formula I

Fertilizer Use Efficiency—

To analyze whether the transgenic plants are more responsive tofertilizers, plants are grown in agar plates or pots with a limitedamount of fertilizer, as described, for example, in Example 6,hereinbelow and in Yanagisawa et al (Proc Natl Acad Sci USA. 2004;101:7833-8). The plants are analyzed for their overall size, time toflowering, yield, protein content of shoot and/or grain. The parameterschecked are the overall size of the mature plant, its wet and dryweight, the weight of the seeds yielded, the average seed size and thenumber of seeds produced per plant. Other parameters that may be testedare: the chlorophyll content of leaves (as nitrogen plant status and thedegree of leaf verdure is highly correlated), amino acid and the totalprotein content of the seeds or other plant parts such as leaves orshoots, oil content, etc. Similarly, instead of providing nitrogen atlimiting amounts, phosphate or potassium can be added at increasingconcentrations. Again, the same parameters measured are the same aslisted above. In this way, nitrogen use efficiency (NUE), phosphate useefficiency (PUE) and potassium use efficiency (KUE) are assessed,checking the ability of the transgenic plants to thrive under nutrientrestraining conditions.

Nitrogen Use Efficiency—

To analyze whether the transgenic Arabidopsis plants are more responsiveto nitrogen, plant are grown in 0.75-1.5 mM (nitrogen deficientconditions) or 6-10 mM (optimal nitrogen concentration). Plants areallowed to grow for additional 20 days or until seed production. Theplants are then analyzed for their overall size, time to flowering,yield, protein content of shoot and/or grain/seed production. Theparameters checked can be the overall size of the plant, wet and dryweight, the weight of the seeds yielded, the average seed size and thenumber of seeds produced per plant. Other parameters that may be testedare: the chlorophyll content of leaves (as nitrogen plant status and thedegree of leaf greenness is highly correlated), amino acid and the totalprotein content of the seeds or other plant parts such as leaves orshoots and oil content. Transformed plants not exhibiting substantialphysiological and/or morphological effects, or exhibiting highermeasured parameters levels than wild-type plants, are identified asnitrogen use efficient plants.

Nitrogen Use Efficiency Assay Using Plantlets—

The assay is done according to Yanagisawa-S. et al. with minormodifications (“Metabolic engineering with Dof1 transcription factor inplants: Improved nitrogen assimilation and growth under low-nitrogenconditions” Proc. Natl. Acad. Sci. USA 101, 7833-7838). Briefly,transgenic plants which are grown for 7-10 days in 0.5×MS[Murashige-Skoog] supplemented with a selection agent are transferred totwo nitrogen-limiting conditions: MS media in which the combinednitrogen concentration (NH₄NO₃ and KNO₃) was 0.2 mM or 0.05 mM. Plantsare allowed to grow for additional 30-40 days and then photographed,individually removed from the Agar (the shoot without the roots) andimmediately weighed (fresh weight) for later statistical analysis.Constructs for which only T1 seeds are available are sown on selectivemedia and at least 25 seedlings (each one representing an independenttransformation event) are carefully transferred to the nitrogen-limitingmedia. For constructs for which T2 seeds are available, differenttransformation events are analyzed. Usually, 25 randomly selected plantsfrom each event are transferred to the nitrogen-limiting media allowedto grow for 3-4 additional weeks and individually weighed at the end ofthat period. Transgenic plants are compared to control plants grown inparallel under the same conditions. Mock-transgenic plants expressingthe uidA reporter gene (GUS) under the same promoter are used ascontrol.

Nitrogen Determination—

The procedure for N (nitrogen) concentration determination in thestructural parts of the plants involves the potassium persulfatedigestion method to convert organic N to NO₃ ⁻ (Purcell and King 1996Argon. J. 88:111-113, the modified Cd⁻ mediated reduction of NO₃ ⁻ toNO₂ ⁻ (Vodovotz 1996 Biotechniques 20:390-394) and the measurement ofnitrite by the Griess assay (Vodovotz 1996, supra). The absorbancevalues are measured at 550 nm against a standard curve of NaNO₂. Theprocedure is described in details in Samonte et al. 2006 Agron. J.98:168-176.

Germination Tests—

Germination tests compare the percentage of seeds from transgenic plantsthat could complete the germination process to the percentage of seedsfrom control plants that are treated in the same manner. Normalconditions are considered for example, incubations at 22° C. under22-hour light 2-hour dark daily cycles. Evaluation of germination andseedling vigor is conducted between 4 and 14 days after planting. Thebasal media is 50% MS medium (Murashige and Skoog, 1962 Plant Physiology15, 473-497).

Germination is checked also at unfavorable conditions such as cold(incubating at temperatures lower than 10° C. instead of 22° C.) orusing seed inhibition solutions that contain high concentrations of anosmolyte such as sorbitol (at concentrations of 50 mM, 100 mM, 200 mM,300 mM, 500 mM, and up to 1000 mM) or applying increasing concentrationsof salt (of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM NaCl).

The effect of the transgene on plant's vigor, growth rate, biomass,yield and/or oil content can be determined using known methods.

Plant Vigor—

The plant vigor can be calculated by the increase in growth parameterssuch as leaf area, fiber length, rosette diameter, plant fresh weightand the like per time.

Growth Rate—

The growth rate can be measured using digital analysis of growingplants. For example, images of plants growing in greenhouse on plotbasis can be captured every 3 days and the rosette area can becalculated by digital analysis. Rosette area growth is calculated usingthe difference of rosette area between days of sampling divided by thedifference in days between samples.

Evaluation of growth rate can be done by measuring plant biomassproduced, rosette area, leaf size or root length per time (can bemeasured in cm² per day of leaf area).

Relative growth area can be calculated using Formula II.

Relative growth rate area=Regression coefficient of area along timecourse  Formula II:

Thus, the relative growth area rate is in units of 1/day and lengthgrowth rate is in units of 1/day.

Seed Yield—

Evaluation of the seed yield per plant can be done by measuring theamount (weight or size) or quantity (i.e., number) of dry seeds producedand harvested from 8-16 plants and divided by the number of plants.

For example, the total seeds from 8-16 plants can be collected, weightedusing e.g., an analytical balance and the total weight can be divided bythe number of plants. Seed yield per growing area can be calculated inthe same manner while taking into account the growing area given to asingle plant. Increase seed yield per growing area could be achieved byincreasing seed yield per plant, and/or by increasing number of plantscapable of growing in a given area.

In addition, seed yield can be determined via the weight of 1000 seeds.The weight of 1000 seeds can be determined as follows: seeds arescattered on a glass tray and a picture is taken. Each sample isweighted and then using the digital analysis, the number of seeds ineach sample is calculated.

The 1000 seeds weight can be calculated using formula III:

1000 Seed Weight=number of seed in sample/sample weight X 1000  FormulaIII:

The Harvest Index can be calculated using Formula IV

Harvest Index=Average seed yield per plant/Average dry weight  FormulaIV:

Grain Protein Concentration—

Grain protein content (g grain protein m⁻²) is estimated as the productof the mass of grain N (g grain N m⁻²) multiplied by the N/proteinconversion ratio of k-5.13 (Mosse 1990, supra). The grain proteinconcentration is estimated as the ratio of grain protein content perunit mass of the grain (g grain protein kg⁻¹ grain).

Fiber Length—

Fiber length can be measured using fibrograph. The fibrograph system wasused to compute length in terms of “Upper Half Mean” length. The upperhalf mean (UHM) is the average length of longer half of the fiberdistribution. The fibrograph measures length in span lengths at a givenpercentage point (Hypertext Transfer Protocol://WorldWide Web (dot)cottoninc (dot) com/ClassificationofCotton/?Pg=4#Length).

According to some embodiments of the invention, increased yield of cornmay be manifested as one or more of the following: increase in thenumber of plants per growing area, increase in the number of ears perplant, increase in the number of rows per ear, number of kernels per earrow, kernel weight, thousand kernel weight (1000-weight), earlength/diameter, increase oil content per kernel and increase starchcontent per kernel.

As mentioned, the increase of plant yield can be determined by variousparameters. For example, increased yield of rice may be manifested by anincrease in one or more of the following: number of plants per growingarea, number of panicles per plant, number of spikelets per panicle,number of flowers per panicle, increase in the seed filling rate,increase in thousand kernel weight (1000-weight), increase oil contentper seed, increase starch content per seed, among others. An increase inyield may also result in modified architecture, or may occur because ofmodified architecture.

Similarly, increased yield of soybean may be manifested by an increasein one or more of the following: number of plants per growing area,number of pods per plant, number of seeds per pod, increase in the seedfilling rate, increase in thousand seed weight (1000-weight), reduce podshattering, increase oil content per seed, increase protein content perseed, among others. An increase in yield may also result in modifiedarchitecture, or may occur because of modified architecture.

Increased yield of canola may be manifested by an increase in one ormore of the following: number of plants per growing area, number of podsper plant, number of seeds per pod, increase in the seed filling rate,increase in thousand seed weight (1000-weight), reduce pod shattering,increase oil content per seed, among others. An increase in yield mayalso result in modified architecture, or may occur because of modifiedarchitecture.

Increased yield of cotton may be manifested by an increase in one ormore of the following: number of plants per growing area, number ofbolls per plant, number of seeds per boll, increase in the seed fillingrate, increase in thousand seed weight (1000-weight), increase oilcontent per seed, improve fiber length, fiber strength, among others. Anincrease in yield may also result in modified architecture, or may occurbecause of modified architecture.

Oil Content—

The oil content of a plant can be determined by extraction of the oilfrom the seed or the vegetative portion of the plant. Briefly, lipids(oil) can be removed from the plant (e.g., seed) by grinding the planttissue in the presence of specific solvents (e.g., hexane or petroleumether) and extracting the oil in a continuous extractor. Indirect oilcontent analysis can be carried out using various known methods such asNuclear Magnetic Resonance (NMR) Spectroscopy, which measures theresonance energy absorbed by hydrogen atoms in the liquid state of thesample [See for example, Conway T F. and Earle F R., 1963, Journal ofthe American Oil Chemists' Society; Springer Berlin/Heidelberg, ISSN:0003-021X (Print) 1558-9331 (Online)]; the Near Infrared (NI)Spectroscopy, which utilizes the absorption of near infrared energy(1100-2500 nm) by the sample; and a method described in WO/2001/023884,which is based on extracting oil a solvent, evaporating the solvent in agas stream which forms oil particles, and directing a light into the gasstream and oil particles which forms a detectable reflected light.

Thus, the present invention is of high agricultural value for promotingthe yield of commercially desired crops (e.g., biomass of vegetativeorgan such as poplar wood, or reproductive organ such as number of seedsor seed biomass).

Any of the transgenic plants described hereinabove or parts thereof maybe processed to produce a feed, meal, protein or oil preparation, suchas for ruminant animals.

The transgenic plants described hereinabove, which exhibit an increasedoil content can be used to produce plant oil (by extracting the oil fromthe plant).

The plant oil (including the seed oil and/or the vegetative portion oil)produced according to the method of the invention may be combined with avariety of other ingredients. The specific ingredients included in aproduct are determined according to the intended use. Exemplary productsinclude animal feed, raw material for chemical modification,biodegradable plastic, blended food product, edible oil, biofuel,cooking oil, lubricant, biodiesel, snack food, cosmetics, andfermentation process raw material. Exemplary products to be incorporatedto the plant oil include animal feeds, human food products such asextruded snack foods, breads, as a food binding agent, aquaculturefeeds, fermentable mixtures, food supplements, sport drinks, nutritionalfood bars, multi-vitamin supplements, diet drinks, and cereal foods.

According to some embodiments of the invention, the oil comprises a seedoil.

According to some embodiments of the invention, the oil comprises avegetative portion oil.

According to some embodiments of the invention, the plant cell forms apart of a plant.

As used herein the term “about” refers to ±10%.

The terms “comprises”, “comprising”, “includes”, “including”, “having”and their conjugates mean “including but not limited to”.

The term “consisting of means “including and limited to”.

The term “consisting essentially of” means that the composition, methodor structure may include additional ingredients, steps and/or parts, butonly if the additional ingredients, steps and/or parts do not materiallyalter the basic and novel characteristics of the claimed composition,method or structure.

As used herein, the singular form “a”, “an” and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention maybe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 3, 4, 5, and 6. This appliesregardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to includeany cited numeral (fractional or integral) within the indicated range.The phrases “ranging/ranges between” a first indicate number and asecond indicate number and “ranging/ranges from” a first indicate number“to” a second indicate number are used herein interchangeably and aremeant to include the first and second indicated numbers and all thefractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniquesand procedures for accomplishing a given task including, but not limitedto, those manners, means, techniques and procedures either known to, orreadily developed from known manners, means, techniques and proceduresby practitioners of the chemical, pharmacological, biological,biochemical and medical arts.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination or as suitable in any other describedembodiment of the invention. Certain features described in the contextof various embodiments are not to be considered essential features ofthose embodiments, unless the embodiment is inoperative without thoseelements.

Various embodiments and aspects of the present invention as delineatedhereinabove and as claimed in the claims section below find experimentalsupport in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with theabove descriptions illustrate some embodiments of the invention in a nonlimiting fashion.

Generally, the nomenclature used herein and the laboratory proceduresutilized in the present invention include molecular, biochemical,microbiological and recombinant DNA techniques. Such techniques arethoroughly explained in the literature. See, for example, “MolecularCloning: A laboratory Manual” Sambrook et al., (1989); “CurrentProtocols in Molecular Biology” Volumes I-III Ausubel, R. M., ed.(1994); Ausubel et al., “Current Protocols in Molecular Biology”, JohnWiley and Sons, Baltimore, Md. (1989); Perbal, “A Practical Guide toMolecular Cloning”, John Wiley & Sons, New York (1988); Watson et al.,“Recombinant DNA”, Scientific American Books, New York; Birren et al.(eds) “Genome Analysis: A Laboratory Manual Series”, Vols. 1-4, ColdSpring Harbor Laboratory Press, New York (1998); methodologies as setforth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and5,272,057; “Cell Biology: A Laboratory Handbook”, Volumes I-III Cellis,J. E., ed. (1994); “Current Protocols in Immunology” Volumes I-IIIColigan J. E., ed. (1994); Stites et al. (eds), “Basic and ClinicalImmunology” (8th Edition), Appleton & Lange, Norwalk, Conn. (1994);Mishell and Shiigi (eds), “Selected Methods in Cellular Immunology”, W.H. Freeman and Co., New York (1980); available immunoassays areextensively described in the patent and scientific literature, see, forexample, U.S. Pat. Nos. 3,791,932; 3,839,153; 3,850,752; 3,850,578;3,853,987; 3,867,517; 3,879,262; 3,901,654; 3,935,074; 3,984,533;3,996,345; 4,034,074; 4,098,876; 4,879,219; 5,011,771 and 5,281,521;“Oligonucleotide Synthesis” Gait, M. J., ed. (1984); “Nucleic AcidHybridization” Hames, B. D., and Higgins S. J., eds. (1985);“Transcription and Translation” Hames, B. D., and Higgins S. J., Eds.(1984); “Animal Cell Culture” Freshney, R. I., ed. (1986); “ImmobilizedCells and Enzymes” IRL Press, (1986); “A Practical Guide to MolecularCloning” Perbal, B., (1984) and “Methods in Enzymology” Vol. 1-317,Academic Press; “PCR Protocols: A Guide To Methods And Applications”,Academic Press, San Diego, Calif. (1990); Marshak et al., “Strategiesfor Protein Purification and Characterization—A Laboratory CourseManual” CSHL Press (1996); all of which are incorporated by reference asif fully set forth herein. Other general references are providedthroughout this document. The procedures therein are believed to be wellknown in the art and are provided for the convenience of the reader. Allthe information contained therein is incorporated herein by reference.

Example 1 Identifying Putative Genes which Increase Abiotic StressTolerance, Yield, Biomass, Growth Rate and/or Fiber Development andQuality

The present inventors have identified genes which increase abioticstress-tolerance (ABST), growth rate, biomass, fiber development orquality, vigor, yield (e.g., seed yield, oil yield), oil content, andnitrogen use efficiency. All nucleotide sequence datasets used here wereoriginated from publicly available databases. Sequence data from 80different plant species was introduced into a single, comprehensivedatabase. Other information on gene expression, protein annotation,enzymes and pathways were also incorporated. Major databases usedinclude:

Genomes

Arabidopsis genome [TAIR genome version 6 (Hypertext TransferProtocol://World Wide Web (dot) arabidopsis (dot) org/)]

Rice genome [IRGSP build 4.0 (Hypertext Transfer Protocol://rgp (dot)dna (dot) affrc (dot) go (dot) jp/IRGSP/)].

Poplar [Populus trichocarpa release 1.1 from JGI (assembly release v1.0)(Hypertext Transfer Protocol://World Wide Web (dot) genome (dot) jgi-psf(dot) org/)]

Brachypodium [JGI 4× assembly Hypertext Transfer Protocol://World WideWeb (dot) brachpodium (dot) org)]

Soybean [DOE-JGI SCP, version Glyma0 (Hypertext TransferProtocol://World Wide Web (dot) phytozome (dot) net/)]

Grape International Grape Genome Program Genome Assembly (HypertextTransfer Protocol://World Wide Web (dot) genoscope (dot) cns (dot)fr/externe/Download/Projets/Projet_mL/data/assembly/

Castobean [TIGR/J Craig Venter Institute 4× assembly (Hypertext TransferProtocol://msc (dot) jcv (dot) org/)]

Sorghum [DOE-JGI SCP, version Sbi1 Hypertext Transfer Protocol://WorldWide Web (dot) phytozome (dot) net/)].

Partially assembled genome of Maize [Hypertext TransferProtocol://maizesequence (dot) org/]

Expressed EST and mRNA sequences were extracted from

GeneBank versions 154, 157, 160, 161, 164, 165, 166 (Hypertext TransferProtocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot)gov/dbEST/)

RefSeq (Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot)nlm (dot) nih (dot) gov/RefSeq/).

TAIR (Hypertext Transfer Protocol://World Wide Web (dot) arabidopsis(dot) org/).

Protein and pathway databases

Uniprot (Hypertext Transfer Protocol://World WideWeb.expasy.uniprot.org/).

AraCyc (Hypertext Transfer Protocol://World Wide Web (dot) arabidopsis(dot) org/biocyc/index (dot) jsp).

ENZYME (Hypertext Transfer Protocol://expasy.org/enzyme/).

Microarray datasets were downloaded from

GEO (Hypertext Transfer Protocol://World Wide Web.ncbi.nlm.nih.gov/geo/)

TAIR (Hypertext Transfer Protocol://World Wide Web.arabidopsis.org/).

Proprietary cotton fiber microarray data

QTL information

Gramene (Hypertext Transfer Protocol://World Wide Web (dot) gramene(dot) org/qtl/).

Database Assembly was performed to build a wide, rich, reliableannotated and easy to analyze database comprised of publicly availablegenomic mRNA, ESTs DNA sequences, data from various crops as well asgene expression, protein annotation and pathway data QTLs, and otherrelevant information.

Database assembly is comprised of a toolbox of gene refining,structuring, annotation and analysis tools enabling to construct atailored database for each gene discovery project. Gene refining andstructuring tools enable to reliably detect splice variants andantisense transcripts, generating understanding of various potentialphenotypic outcomes of a single gene. The capabilities of the “LEADS”platform of Compugen LTD for analyzing human genome have been confirmedand accepted by the scientific community (“Widespread AntisenseTranscription”, Yelin, et al. (2003) Nature Biotechnology 21, 379-85;“Splicing of Alu Sequences”, Lev-Maor, et al. (2003) Science 300 (5623),1288-91; “Computational analysis of alternative splicing using ESTtissue information”, Xie H et al. Genomics 2002), and have been provenmost efficient in plant genomics as well.

EST Clustering and Gene Assembly—

For gene clustering and assembly of organisms with available genomesequence data (arabidopsis, rice, castorbean, grape, brachypodium,poplar, soybean, sorghum) the genomic LEADS version (GANG) was employed.This tool allows most accurate clustering of ESTs and mRNA sequences ongenome, and predicts gene structure as well as alternative splicingevents and anti-sense transcription.

For organisms with no available full genome sequence data, “expressedLEADS” clustering software was applied.

Gene Annotation—

Predicted genes and proteins were annotated as follows:

Blast search (Hypertext Transfer Protocol://World Wide Web (dot) ncbi(dot) nlm (dot) nih (dot) gov (dot) library (dot) vu (dot) edu (dot)au/BLAST/) against all plant UniProt (Hypertext TransferProtocol://World Wide Web (dot) expasy (dot) uniprot (dot) org/)sequences was performed.

Open reading frames of each putative transcript were analyzed andlongest ORF with higher number of homologs was selected as predictedprotein of the transcript.

The predicted proteins were analyzed by InterPro (Hypertext TransferProtocol://World Wide Web (dot) ebi (dot) ac (dot) uk/interpro/).

Blast against proteins from AraCyc and ENZYME databases was used to mapthe predicted transcripts to AraCyc pathways.

Predicted proteins from different species were compared using blastalgorithm (Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot)nlm (dot) nih (dot) gov (dot) library (dot) vu (dot) edu (dot)au/BLAST/) to validate the accuracy of the predicted protein sequence,and for efficient detection of orthologs.

Gene Expression Profiling—

Few data sources were exploited for gene expression profiling, namelymicroarray data and digital expression profile (see below). According togene expression profile, a correlation analysis was performed toidentify genes which are co-regulated under different development stagesand environmental conditions.

Publicly available microarray datasets were downloaded from TAIR andNCBI GEO sites, renormalized, and integrated into the database.Expression profiling is one of the most important resource data foridentifying genes important for ABST. Moreover, when homolog genes fromdifferent crops were responsive to ABST, the genes are marked as “highlypredictive to improve ABST”.

A digital expression profile summary was compiled for each clusteraccording to all keywords included in the sequence records comprisingthe cluster. Digital expression, also known as electronic Northern Blot,is a tool that displays virtual expression profile based on the ESTsequences forming the gene cluster. The tool can provide the expressionprofile of a cluster in terms of plant anatomy (in what tissues/organsis the gene expressed), developmental stage (the developmental stages atwhich a gene can be found) and profile of treatment (provides thephysiological conditions under which a gene is expressed such asdrought, cold, pathogen infection, etc). Given a random distribution ofESTs in the different clusters, the digital expression provides aprobability value that describes the probability of a cluster having atotal of N ESTs to contain X ESTs from a certain collection oflibraries. For the probability calculations are taken intoconsideration: a) the number of ESTs in the cluster, b) the number ofESTs of the implicated and related libraries, c) the overall number ofESTs available representing the species. Thereby clusters with lowprobability values are highly enriched with ESTs from the group oflibraries of interest indicating a specialized expression.

The results of the digital and microarray gene expression data areprovided in Tables 1-4, hereinbelow.

Below are summarized the key criteria used to select the genes whichexpression thereof in a plant can be used to increase ABST, WUE, NUE,FUE, biomass, yield and oil content. The overexpression Fold (“Fold”) iscalculated as the ratio between the number of ESTs found in a gene or anorthologue group for a certain category (“Keyword”) and the number ofexpected ESTs according to a normal distribution. A probabilistic value(P-value) was estimated for the calculated overexpression folds. Geneswere selected based on the results presented in Tables 1-4 below andother computational filtering combined with manual curation as detailedbelow.

LAB25, LAB31, LAB33, LAB34, LAB45 and LAB51 were selected since they arehighly expressed in roots and under drought stress conditions (as shownin Table 1 hereinbelow).

TABLE 1 Digital expression of LAB25, LAB31, LAB33, LAB34, LAB45 andLAB51 in roots and under drought stress Anatomy Treatment Root Droughtstress Genes fold p-value fold p-value LAB25 5.39 1.125E−52 1.9844340.0404051 LAB31 10.00 6.034E−09 7.00 8.6157E−06 LAB33 2.66 7.272E−053.25  0.00090165 LAB34 3.38 1.474E−05 9.55 6.8734E−08 LAB45 2.22 1.7E−07 14.11 4.2333E−14 LAB51 2.10 0.0046312 4.00 0.0131351 Table 1.Digital expression of the indicated genes in root and under droughtstress. Provided are the fold increase and the calculated p-values ofexpression of the gene in the indicated tissue or condition as comparedto the randomly expected expression. Results were consideredstatistically significant if the p-value was lower than 0.05.

LAB4, LAB7, LAB14 and LAB49 were selected since they are highlyexpressed in roots and under UV radiation, cold stress or heat stress(as shown in Table 2 hereinbelow).

TABLE 2 Digital expression of LAB4, LAB7, LAB14 and LAB49 in roots,under UV irradiation, cold stress or heat stress Anatomy Treatment RootUV irradiation Cold stress Heat stress Genes fold p-value fold p-valuefold p-value fold p-value LAB4 4.45 2.005E−10 LAB7 2.48 6.421E−08 2.370.0303 LAB14 2.15 0.0319954 3.64 0.00019 2 0.0570 LAB49 4.17 8.6877E−11 Table 2. Digital expression of the indicated genes in roots, under UVirradiation, cold stress or heat stress. Provided are the fold increaseand the calculated p-values of expression of the gene in the indicatedtissue or condition as compared to the randomly expected expression.Results were considered statistically significant if the p-value waslower than 0.05. Blank cells indicate that either the gene is notexpressed or data is not available.

LAB5, LAB13, LAB16, LAB18, LAB20, LAB22, LAB3, LAB24, LAB35, LAB38,LAB39, LAB40, LAB50 and LAB51 were selected since they are highlyexpressed under drought stress and possibly nutrient deficiencies, coldstress or plant development or stress hormones (as shown in Table 3hereinbelow).

TABLE 3 Digital expression of LAB5, LAB13, LAB16, LAB18, LAB20, LAB22,LAB3, LAB24, LAB35, LAB38, LAB39, LAB40, LAB50 and LAB51 under droughtstress and possibly nutrient deficiencies, cold stress or plantdevelopment or stress hormones Nutrient deficiencies Plant developmentDrought stress Cold or stress fold p-value stress hormones 3.460.00188373 fold p-value fold p-value fold p-value LAB5 3.13 0.0400183LAB13 3.00 0.00017491 LAB16 4.00 0.00458478 LAB18 4.95 4.2144E−05 LAB208.88 3.4638E−22 LAB22 3.00 0.00978408 3.17 0.0379553 LAB3 2.375.7818E−08 LAB24 14.11 4.2333E−14 LAB35 4.00 0.00207373 3.00 0.0072537LAB38 2.35 0.00067594 LAB39 8.93 2.6849E−08 3.06 0.0144515 LAB40 7.005.6733E−05 LAB50 3.44 1.1207E−06 3.15 0.012142 Table 3. Digitalexpression of the indicated genes under drought stress, possiblynutrient deficiencies, cold stress or plant development or stresshormones. Provided are the fold increase and the calculated p-values ofexpression of the gene in the indicated tissue or condition as comparedto the randomly expected expression. Results were consideredstatistically significant if the p-value was lower than 0.05. Blankcells indicate that either the gene is not expressed or data is notavailable.

LAB9, LAB21, LAB32, LAB15, LAB17, LAB30, LAB36, and LAB39 were selectedsince they are highly expressed under etiolatlion condition, plantdevelopment or stress hormones, salinity stress or waterlogging (asshown in Table 4 hereinbelow).

TABLE 4 Digital expression of LAB9, LAB21, LAB32, LAB15, LAB17, LAB30,LAB36, and LAB39 under etiolatlion condition, plant development orstress hormones, salinity stress or waterlogging Etiolated Plantdevelopment or stress hormones fold p-value Salinity stress Waterlogging2.91 0.0160756 fold p-value fold p-value fold p-value LAB9 2.230.00043618 LAB21 4.65 5.5967E−17 LAB32 LAB15 1.0 0.0705542 LAB17 2.00.0420927 LAB30 6.00 7.4196E−05 LAB36 3.66  3.338E−06 4.7 9.3682E−06LAB39 Table 4. Digital expression of the indicated genes underetiolatlion condition, plant development or stress hormones, salinitystress or waterlogging. Provided are the fold increase and thecalculated p-values of expression of the gene in the indicated tissue orcondition as compared to the randomly expected expression. Results wereconsidered statistically significant if the p-value was lower than 0.05.Blank cells indicate that either the gene is not expressed or data isnot available.

Overall, 51 genes were identified to have a major impact on ABST,nitrogen use efficiency, yield (e.g., seed yield), oil content, growthrate and/or vigor when overexpressed in plants. The identified genes,their curated polynucleotide and polypeptide sequences, as well as theirupdated sequences according to Genebank database are summarized in Table5, hereinbelow.

TABLE 5 Identified genes which can be used to increase ABST, fiberdevelopment (quality and yield), yield, biomass, growth rate, nitrogenuse efficiency, fertilizer use efficiency, water use efficiency, and/oroil content of a plant SEQ ID NO: SEQ ID NO: Gene Name Cluster NameOrganism Polynuc. Polypep. LAB4 rice|gb157.2|AA751809 rice 1 60 LAB5sorghum|gb161.xeno|AW922806 sorghum 2 61 LAB7 rice|gb157.2|AA754242 rice3 62 LAB8 rice|gb157.2|AA754407 rice 4 63 LAB9 rice|gb157.2|AB004799rice 5 64 LAB11 rice|gb157.2|AK070868 rice 6 65 LAB13rice|gb157.2|AT003625 rice 7 66 LAB14 rice|gb157.2|AU056017 rice 8 67LAB15 barley|gb157.3|BF623077 barley 9 68 LAB2 barley|gb157.3|BE195266barley 10 69 LAB16 cotton|gb164|BE052656 cotton 11 70 LAB17sorghum|gb161.xeno|AI724026 sorghum 12 71 LAB18sorghum|gb161.xeno|BE359151 sorghum 13 72 LAB20 rice|gb157.2|AW070136rice 14 73 LAB21 barley|gb157.3|BE421259 barley 15 74 LAB22sorghum|gb161.xeno|AW678130 sorghum 16 75 LAB3 canola|gb161|CD831005canola 17 76 LAB23 barley|gb157.3|BI947386 barley 18 77 LAB24sorghum|gb161.xeno|AW433371 sorghum 19 78 LAB25 barley|gb157.3|X84056barley 20 79 LAB30 sorghum|gb161.xeno|BE362140 sorghum 21 80 LAB31canola|gb161|H74460 canola 22 81 LAB32 barley|gb157.3|AL499903 barley 2382 LAB33 sorghum|gb161.xeno|AW676682 sorghum 24 83 LAB34soybean|gb166|CF921741 soybean 25 84 LAB35 wheat|gb164|BE497867 wheat 2685 LAB36 sorghum|gb161.xeno|H55004 sorghum 27 86 LAB38wheat|gb164|BE412185 wheat 28 87 LAB39 sorghum|gb161.xeno|BG048297sorghum 29 88 LAB40 wheat|gb164|BE488436 wheat 30 89 LAB41wheat|gb164|X52472 wheat 31 90 LAB43 barley|gb157.3|BF624177 barley 3291 LAB45 sorghum|gb161.crp|AI855293 sorghum 33 92 LAB49rice|gb157.2|BE040470 rice 34 93 LAB50 rice|gb157.2|BI305323 rice 35 94LAB51 wheat|gb164|BI751966 wheat 36 95 BDL103_P1 rice|gb157.2|BE228840rice 37 96 BDL11 arabidopsis|gb165|AT5G12460 arabidopsis 38 97 BDL12arabidopsis|gb165|AT4G08530 arabidopsis 39 98 BDL14arabidopsis|gb165|AT1G53690 arabidopsis 40 99 BDL166arabidopsis|gb165|AT1G71691 arabidopsis 41 100 BDL17arabidopsis|gb165|AT5G36680 arabidopsis 42 101 BDL210arabidopsis|gb165|AT5G22810 arabidopsis 43 102 CTF113cotton|gb164|AI727515 cotton 44 103 CTF163 cotton|gb164|CO123733 cotton45 104 CTF175 cotton|gb164|AW187393 cotton 46 105 CTF180cotton|gb164|BG440663 cotton 47 106 CTF205 cotton|gb164|AI725800 cotton48 107 CTF215 cotton|gb164|AI729467 cotton 49 108 CTF225cotton|gb164|AW187127 cotton 50 109 CTF226 cotton|gb164|AI730124 cotton51 110 LAB2 barley|gb157.3|BE195266 barley 52 69 LAB3canola|gb161|CD831005 canola 53 76 LAB32 barley|gb157.3|AL499903 barley54 82 LAB38 wheat|gb164|BE412185 wheat 55 87 LAB51 wheat|gb164|BI751966wheat 56 95 BDL17 arabidopsis|gb165|AT5G36680 arabidopsis 57 111 CTF163cotton|gb164|CO123733 cotton 58 104 CTF205 cotton|gb164|AI725800 cotton59 112 BDL103_P2 rice|gb157.2|BE228840 rice 638 96 Table 5. Provided arethe identified genes, their annotation, organism and polynucleotide andpolypeptide sequence identifiers. SEQ ID NOs: 52-59 are polynucleotidesequences which were uncovered after cloning the gene. SEQ ID NO: 638 isa computational curated sequence.

Example 2 Identification of Homologues which Affect ABST, WUE, NUE, FUE,Yield, Growth Rate, Vigor, Biomass and Oil Content

The concepts of orthology and paralogy have been applied to functionalcharacterizations and classifications on the scale of whole-genomecomparisons. Orthologs and paralogs constitute two major types ofhomologs: The first evolved from a common ancestor by specialization,and the latter are related by duplication events. It is assumed thatparalogs arising from ancient duplication events are likely to havediverged in function while true orthologs are more likely to retainidentical function over evolutionary time.

To further investigate and identify putative ortholog genes of genesaffecting abiotic stress tolerance, nitrogen use efficiency, fertilizeruse efficiency, yield (e.g., seed yield, oil yield, biomass, grainquantity and/or quality), growth rate, vigor, biomass, oil content,and/or water use efficiency (presented in Table 5, above) all sequenceswere aligned using the BLAST (/Basic Local Alignment Search Tool/).Sequences sufficiently similar were tentatively grouped. These putativeorthologs were further organized under a Phylogram—a branching diagram(tree) assumed to be a representation of the evolutionary relationshipsamong the biological taxa. Putative ortholog groups were analyzed as totheir agreement with the phylogram and in cases of disagreements theseortholog groups were broken accordingly. Expression data was analyzedand the EST libraries were classified using a fixed vocabulary of customterms such as developmental stages (e.g., genes showing similarexpression profile through development with up regulation at specificstage, such as at the seed filling stage) and/or plant organ (e.g.,genes showing similar expression profile across their organs with upregulation at specific organs such as root). The annotations from allthe ESTs clustered to a gene were analyzed statistically by comparingtheir frequency in the cluster versus their abundance in the database,allowing the construction of a numeric and graphic expression profile ofthat gene, which is termed “digital expression”. The rationale of usingthese two complementary methods with methods of phenotypic associationstudies of QTLs, and phenotype expression correlation is based on theassumption that true orthologs are likely to retain identical functionover evolutionary time. These methods provide different sets ofindications on function similarities between two homologous genes,similarities in the sequence level—identical amino acids in the proteindomains and similarity in expression profiles.

The search and identification of homologous genes involves the screeningof sequence information available, for example, in public databases,which include but are not limited to the DNA Database of Japan (DDBJ),Genbank, and the European Molecular Biology Laboratory Nucleic AcidSequence Database (EMBL) or versions thereof or the MIPS database. Anumber of different search algorithms have been developed, including butnot limited to the suite of programs referred to as BLAST programs.There are five implementations of BLAST, three designed for nucleotidesequence queries (BLASTN, BLASTX, and TBLASTX) and two designed forprotein sequence queries (BLASTP and TBLASTN) (Coulson, Trends inBiotechnology: 76-80, 1994; Birren et al., Genome Analysis, I: 543,1997). Such methods involve alignment and comparison of sequences. TheBLAST algorithm calculates percent sequence identity and performs astatistical analysis of the similarity between the two sequences. Thesoftware for performing BLAST analysis is publicly available through theNational Centre for Biotechnology Information. Other such software oralgorithms are GAP, BESTFIT, FASTA and TFASTA. GAP uses the algorithm ofNeedleman and Wunsch (J. Mol. Biol. 48: 443-453, 1970) to find thealignment of two complete sequences that maximizes the number of matchesand minimizes the number of gaps.

The homologous genes may belong to the same gene family. The analysis ofa gene family may be carried out using sequence similarity analysis. Toperform this analysis one may use standard programs for multiplealignments e.g. Clustal W. A neighbor-joining tree of the proteinshomologous to the genes of some embodiments of the invention may be usedto provide an overview of structural and ancestral relationships.Sequence identity may be calculated using an alignment program asdescribed above. It is expected that other plants will carry a similarfunctional gene (orthologue) or a family of similar genes and thosegenes will provide the same preferred phenotype as the genes presentedhere. Advantageously, these family members may be useful in the methodsof some embodiments of the invention. Example of other plants include,but not limited to, barley (Hordeum vulgare), Arabidopsis (Arabidopsisthaliana), maize (Zea mays), cotton (Gossypium), Oilseed rape (Brassicanapus), Rice (Oryza sativa), Sugar cane (Saccharum officinarum), Sorghum(Sorghum bicolor), Soybean (Glycine max), Sunflower (Helianthus annuus),Tomato (Lycopersicon esculentum) and Wheat (Triticum aestivum).

The above-mentioned analyses for sequence homology is preferably carriedout on a full-length sequence, but may also be based on a comparison ofcertain regions such as conserved domains. The identification of suchdomains would also be well within the realm of the person skilled in theart and would involve, for example, a computer readable format of thenucleic acids of some embodiments of the invention, the use of alignmentsoftware programs and the use of publicly available information onprotein domains, conserved motifs and boxes. This information isavailable in the PRODOM (Hypertext Transfer Protocol://World Wide Web(dot) biochem (dot) ucl (dot) ac (dot)uk/bsm/dbbrowser/protocol/prodomqry (dot) html), PIR (Hypertext TransferProtocol://pir (dot) Georgetown (dot) edu/) or Pfam (Hypertext TransferProtocol://World Wide Web (dot) sanger (dot) ac (dot) uk/Software/Pfam/)database. Sequence analysis programs designed for motif searching may beused for identification of fragments, regions and conserved domains asmentioned above. Preferred computer programs include, but are notlimited to, MEME, SIGNALSCAN, and GENESCAN.

A person skilled in the art may use the homologous sequences providedherein to find similar sequences in other species and other organisms.Homologues of a protein encompass, peptides, oligopeptides,polypeptides, proteins and enzymes having amino acid substitutions,deletions and/or insertions relative to the unmodified protein inquestion and having similar biological and functional activity as theunmodified protein from which they are derived. To produce suchhomologues, amino acids of the protein may be replaced by other aminoacids having similar properties (conservative changes, such as similarhydrophobicity, hydrophilicity, antigenicity, propensity to form orbreak a-helical structures or 3-sheet structures). Conservativesubstitution Tables are well known in the art [see for example Creighton(1984) Proteins. W.H. Freeman and Company]. Homologues of a nucleic acidencompass nucleic acids having nucleotide substitutions, deletionsand/or insertions relative to the unmodified nucleic acid in questionand having similar biological and functional activity as the unmodifiednucleic acid from which they are derived.

Table 6, hereinbelow, lists a summary of orthologous and homologoussequences of the polynucleotide sequences (SEQ ID NOs:1-59 and 638) andpolypeptide sequences (SEQ ID NOs:60-112) presented in Table 5, whichwere identified using NCBI BLAST (BlastP) and needle (EMBOSS package)having at least 80% identity to the selected polypeptides and which areexpected to posses the same role in abiotic stress tolerance (ABST),water use efficiency (WUE), nitrogen use efficiency (NUE), fertilizeruse efficiency (FUE), biomass increment, growth rate increment, yield,vigor, fiber quality and/or yield and/or oil content of plants.

TABLE 6 Homologues of the identified genes of the invention which canincrease ABST, fiber development (quality and yield), biomass, growthrate, nitrogen use efficiency, fertilizer use efficiency, water useefficiency, yield and/or oil content of a plant Polynucl. Polypep.Homology SEQ ID Gene SEQ ID to SEQ % Global NO: Name Organism/Clustername NO: ID NO: identity Algor. 113 LAB4_H0 sorghum|gb161.crp|AW747731359 60 82.6 blastp 114 LAB4_H1 switchgrass|gb167|DN143443 360 60 81.6blastp 115 LAB5_H0 barley|gb157.3|BE412466 361 61 83.1 blastp 116LAB5_H1 barley|gb157.3|BF623020 362 61 82.3 blastp 117 LAB5_H2barley|gb157.3|BI953964 363 61 82.3 blastp 118 LAB5_H3brachypodium|gb169|AF181661 364 61 85.61 tblastn 119 LAB5_H4cenchrus|gb166|BM084156 365 61 85.1 blastp 120 LAB5_H5fescue|gb161|DT683694 366 61 87.7 blastp 121 LAB5_H6maize|gb170|BI325281 367 61 94.7 blastp 122 LAB5_H7rice|gb170|OS06G46950 368 61 82.3 blastp 123 LAB5_H8 rye|gb164|BE637379369 61 80 tblastn 124 LAB5_H9 spruce|gb162|CO219921 370 61 83.1 blastp125 LAB5_H10 sugarcane|gb157.3|BQ529602 371 61 93.3 blastp 126 LAB5_H11sugarcane|gb157.3|BQ535202 372 61 89.23 tblastn 127 LAB5_H12sugarcane|gb157.3|CA072503 373 61 94.8 blastp 128 LAB5_H13sugarcane|gb157.3|CA082920 374 61 94.1 blastp 129 LAB5_H14sugarcane|gb157.3|CA085102 375 61 89.1 blastp 130 LAB5_H15sugarcane|gb157.3|CA090891 376 61 94.1 blastp 131 LAB5_H16sugarcane|gb157.3|CA122790 377 61 90.5 blastp 132 LAB5_H17switchgrass|gb167|DN145030 378 61 80.6 blastp 133 LAB5_H18switchgrass|gb167|FE635988 379 61 89.3 blastp 134 LAB5_H19switchgrass|gb167|FL774816 380 61 85.5 tblastn 135 LAB5_H20wheat|gb164|AF181661 381 61 84.6 blastp 136 LAB5_H21wheat|gb164|BE417364 382 61 83.1 blastp 137 LAB5_H22wheat|gb164|BF484215 383 61 80.6 blastp 138 LAB7_H0barley|gb157.3|AL501769 384 62 83 blastp 139 LAB7_H1brachypodium|gb169|BE471170 385 62 82.8 blastp 140 LAB7_H2maize|gb170|AW042403 386 62 80.4 blastp 141 LAB7_H3 maize|gb170|T69041387 62 81.8 blastp 142 LAB7_H4 sorghum|gb161.crp|BE356561 388 62 82.3blastp 143 LAB7_H5 sugarcane|gb157.3|CA091573 389 62 83.1 blastp 144LAB7_H6 switchgrass|gb167|DN142661 390 62 81.4 blastp 145 LAB7_H7switchgrass|gb167|FE615102 391 62 81.2 blastp 146 LAB7_H8wheat|gb164|BE443254 392 62 84 blastp 147 LAB7_H9 wheat|gb164|BE471170393 62 83.1 blastp 148 LAB7_H10 wheat|gb164|BF293813 394 62 82.6 blastp149 LAB8_H0 rice|gb170|OS03G22790 395 63 98.85 tblastn 150 LAB15_H0wheat|gb164|BM137033 396 68 87.63 tblastn 151 LAB15_H1wheat|gb164|BM138703 397 68 83.51 tblastn 152 LAB15_H2wheat|gb164|CD882022 398 68 81.5 blastp 153 LAB2_H0brachypodium|gb169|DV485170 399 69 81.8 blastp 154 LAB2_H1fescue|gb161|DT694419 400 69 83.2 blastp 155 LAB16_H0antirrhinum|gb166|AJ787590 401 70 81.2 blastp 156 LAB16_H1apple|gb171|CN580957 402 70 86.4 blastp 157 LAB16_H2apricot|gb157.2|CB824020 403 70 85.1 blastp 158 LAB16_H3arabidopsis|gb165|AT4G38580 404 70 83.7 blastp 159 LAB16_H4b_juncea|gb164|EVGN00544315151807 405 70 83.7 blastp 160 LAB16_H5b_oleracea|gb161|AM058105 406 70 82.4 blastp 161 LAB16_H6b_oleracea|gb161|ES942384 407 70 81.7 tblastn 162 LAB16_H7b_rapa|gb162|EX025293 408 70 83 blastp 163 LAB16_H8barley|gb157.3|BF258224 409 70 84.97 tblastn 164 LAB16_H9bean|gb167|CA910356 410 70 85.1 blastp 165 LAB16_H10cacao|gb167|CU476614 411 70 95.5 blastp 166 LAB16_H11canola|gb161|CD817401 412 70 83 blastp 167 LAB16_H12canola|gb161|CN736951 413 70 81.7 blastp 168 LAB16_H13cassava|gb164|BI325222 414 70 84.42 tblastn 169 LAB16_H14castorbean|09v1|EG691829 415 70 87 blastp 170 LAB16_H15catharanthus|gb166|FD421293 416 70 84.4 blastp 171 LAB16_H16chestnut|gb170|SRR006295S0010879 417 70 81.3 blastp 172 LAB16_H17chickpea|09v1|FE673275 418 70 81.4 blastp 173 LAB16_H18cichorium|gb171|EH697988 419 70 80 blastp 174 LAB16_H19citrus|gb166|CN184469 420 70 80.5 blastp 175 LAB16_H20coffea|gb157.2|DV666808 421 70 80.6 blastp 176 LAB16_H21cowpea|gb166|FC458156 422 70 83.1 blastp 177 LAB16_H22cowpea|gb166|FF538669 423 70 80.8 blastp 178 LAB16_H23grape|gb160|BM436505 424 70 86.5 blastp 179 LAB16_H24ipomoea|gb157.2|BJ555808 425 70 82.6 blastp 180 LAB16_H25kiwi|gb166|FG420453 426 70 81.8 blastp 181 LAB16_H26liquorice|gb171|FS257949 427 70 80.5 blastp 182 LAB16_H27lotus|09v1|GO007127 428 70 84.6 blastp 183 LAB16_H28medicago|09v1|BE320877 429 70 81.4 blastp 184 LAB16_H29melon|gb165|AM726967 430 70 83.1 blastp 185 LAB16_H30papaya|gb165|EX255354 431 70 86.5 blastp 186 LAB16_H31peach|gb157.2|BU039481 432 70 86.4 blastp 187 LAB16_H32peanut|gb171|CX018165 433 70 82.2 blastp 188 LAB16_H33peanut|gb171|ES491048 434 70 82.8 blastp 189 LAB16_H34pepper|gb171|BM060814 435 70 81.8 blastp 190 LAB16_H35periwinkle|gb164|FD421293 436 70 84.4 blastp 191 LAB16_H36poplar|gb170|AJ534494 437 70 85.1 blastp 192 LAB16_H37poplar|gb170|BI129301 438 70 83.8 blastp 193 LAB16_H38potato|gb157.2|BG098018 439 70 82.5 blastp 194 LAB16_H39potato|gb157.2|BG098308 440 70 82.5 blastp 195 LAB16_H40prunus|gb167|BU039481 441 70 86.4 blastp 196 LAB16_H41radish|gb164|EV544328 442 70 83.7 blastp 197 LAB16_H42soybean|gb168|BE315834 443 70 85.1 blastp 198 LAB16_H43spurge|gb161|DV122649 444 70 85.71 tblastn 199 LAB16_H44strawberry|gb164|CO817272 445 70 83.8 blastp 200 LAB16_H45tomato|gb164|AA824901 446 70 81.8 blastp 201 LAB16_H46triphysaria|gb164|EX989778 447 70 81.8 blastp 202 LAB16_H47walnuts|gb166|CB303653 448 70 80.5 blastp 203 LAB17_H0barley|gb157.3|BE231003 449 71 91 blastp 204 LAB17_H1brachypodium|gb169|BE498333 450 71 91 blastp 205 LAB17_H2cenchrus|gb166|EB657534 451 71 84.5 blastp 206 LAB17_H3fescue|gb161|DT685866 452 71 91.6 blastp 207 LAB17_H4leymus|gb166|EG394438 453 71 90.3 blastp 208 LAB17_H5maize|gb170|AW498181 454 71 94.2 blastp 209 LAB17_H6pseudoroegneria|gb167|FF340520 455 71 90.3 blastp 210 LAB17_H7rice|gb170|OS04G17100 456 71 93.5 blastp 211 LAB17_H8sugarcane|gb157.3|CA073067 457 71 85.16 tblastn 212 LAB17_H9sugarcane|gb157.3|CA075729 458 71 96.8 blastp 213 LAB17_H10sugarcane|gb157.3|CA078804 459 71 96.8 blastp 214 LAB17_H11sugarcane|gb157.3|CA116673 460 71 96.1 blastp 215 LAB17_H12sugarcane|gb157.3|CA118688 461 71 97.4 blastp 216 LAB17_H13sugarcane|gb157.3|CA119291 462 71 96.1 blastp 217 LAB17_H14sugarcane|gb157.3|CA222723 463 71 94.2 blastp 218 LAB17_H15switchgrass|gb167|DN143094 464 71 85.9 blastp 219 LAB17_H16switchgrass|gb167|FL792168 465 71 80.8 blastp 220 LAB17_H17wheat|gb164|BE498333 466 71 90.3 blastp 221 LAB17_H18wheat|gb164|BF474623 467 71 89.7 blastp 222 LAB17_H19wheat|gb164|CV760043 468 71 89.7 blastp 223 LAB18_H0switchgrass|gb167|DN140747 469 72 80.6 blastp 224 LAB20_H0sugarcane|gb157.3|CA130714 470 73 82.6 blastp 225 LAB21_H0aquilegia|gb157.3|DR914842 471 74 82.9 blastp 226 LAB21_H1arabidopsis|gb165|AT3G47340 472 74 80.6 blastp 227 LAB21_H2b_oleracea|gb161|X84448 473 74 80.4 blastp 228 LAB21_H3b_rapa|gb162|CV545962 474 74 80.3 blastp 229 LAB21_H4barley|gb157.3|BI948886 475 74 87.5 blastp 230 LAB21_H5bean|gb167|AJ133522 476 74 81.9 blastp 231 LAB21_H6 bean|gb167|CB542570477 74 83.1 blastp 232 LAB21_H7 cacao|gb167|CA797951 478 74 83.5 blastp233 LAB21_H8 castorbean|09v1|EE256522 479 74 82.1 blastp 234 LAB21_H9centaurea|gb166|EL931554 480 74 81.2 blastp 235 LAB21_H10citrus|gb166|BQ623162 481 74 83 blastp 236 LAB21_H11cotton|gb164|AI054642 482 74 83.1 blastp 237 LAB21_H12cotton|gb164|BF277939 483 74 81.8 blastp 238 LAB21_H13cotton|gb164|CD486005 484 74 84 blastp 239 LAB21_H14cowpea|gb166|FC458174 485 74 83.3 blastp 240 LAB21_H15cowpea|gb166|FC461749 486 74 82.8 blastp 241 LAB21_H16kiwi|gb166|FG404880 487 74 81.8 blastp 242 LAB21_H17lettuce|gb157.2|DW062326 488 74 83.1 blastp 243 LAB21_H18maize|gb170|AW076472 489 74 85.1 blastp 244 LAB21_H19medicago|09v1|AW126175 490 74 80.7 blastp 245 LAB21_H20monkeyflower|09v1|GO982561 491 74 81.4 blastp 246 LAB21_H21oak|gb170|CU656355 492 74 82.8 blastp 247 LAB21_H22oil_palm|gb166|EL681380 493 74 87 blastp 248 LAB21_H23peach|gb157.2|BU043116 494 74 82 blastp 249 LAB21_H24poplar|gb170|BI138803 495 74 82.8 blastp 250 LAB21_H25poplar|gb170|BU814657 496 74 83.6 blastp 251 LAB21_H26potato|gb157.2|CK258159 497 74 82.1 blastp 252 LAB21_H27prunus|gb167|BU043116 498 74 82.7 blastp 253 LAB21_H28pseudoroegneria|gb167|FF342746 499 74 98.3 blastp 254 LAB21_H29radish|gb164|AB050900 500 74 80.6 blastp 255 LAB21_H30sorghum|gb161.crp|AW286475 501 74 84.96 tblastn 256 LAB21_H31soybean|gb168|AW126284 502 74 83.1 blastp 257 LAB21_H32soybean|gb168|AW720554 503 74 82.3 blastp 258 LAB21_H33soybean|gb168|GMU55874 504 74 81.48 tblastn 259 LAB21_H34soybean|gb168|GMU77678 505 74 83.5 blastp 260 LAB21_H35soybean|gb168|GMU77679 506 74 83 blastp 261 LAB21_H36sugarcane|gb157.3|BQ535363 507 74 85.5 blastp 262 LAB21_H37sugarcane|gb157.3|BQ535939 508 74 85.5 blastp 263 LAB21_H38sunflower|gb162|AF037363 509 74 81.3 blastp 264 LAB21_H39sunflower|gb162|AF190728 510 74 80.9 blastp 265 LAB21_H40sunflower|gb162|DY931765 511 74 80.7 blastp 266 LAB21_H41tomato|gb164|BG127495 512 74 82.5 blastp 267 LAB21_H42triphysaria|gb164|AF014055 513 74 81.9 blastp 268 LAB21_H43wheat|gb164|BE403264 514 74 98.6 blastp 269 LAB21_H44wheat|gb164|BE403866 515 74 98.6 blastp 270 LAB21_H45wheat|gb164|BE430398 516 74 87.9 blastp 271 LAB22_H0maize|gb170|BG833173 517 75 81.7 blastp 272 LAB22_H1maize|gb170|BI423707 518 75 86.4 blastp 273 LAB22_H2sugarcane|gb157.3|BQ536240 519 75 90.9 blastp 274 LAB22_H3sugarcane|gb157.3|BQ536340 520 75 91 blastp 275 LAB22_H4sugarcane|gb157.3|BU103170 521 75 88.2 blastp 276 LAB22_H5sugarcane|gb157.3|CA116439 522 75 89.4 blastp 277 LAB22_H6switchgrass|gb167|FL733549 523 75 85.9 blastp 278 LAB22_H7wheat|gb164|CA484841 524 75 100 blastp 279 LAB3_H0arabidopsis|gb165|AT1G15380 525 76 89.1 blastp 280 LAB3_H1b_rapa|gb162|EX016736 526 76 98.9 blastp 281 LAB3_H2canola|gb161|CD830331 527 76 81.6 blastp 282 LAB3_H3canola|gb161|CN731229 528 76 81.6 blastp 283 LAB3_H4radish|gb164|EV527368 529 76 94.3 blastp 284 LAB3_H5radish|gb164|EV547102 530 76 81 blastp 285 LAB23_H0pseudoroegneria|gb167|FF341473 531 77 89.9 blastp 286 LAB23_H1wheat|gb164|BE516915 532 77 87.6 blastp 287 LAB23_H2wheat|gb164|BE517204 533 77 88.48 tblastn 288 LAB24_H0maize|gb170|BE552559 534 78 80.4 blastp 289 LAB24_H1sugarcane|gb157.3|CA087195 535 78 86.1 blastp 290 LAB24_H2switchgrass|gb167|FL736257 536 78 80 blastp 291 LAB25_H0leymus|gb166|EG374989 537 79 85.6 blastp 292 LAB25_H1leymus|gb166|EG375019 538 79 91.3 blastp 293 LAB25_H2pseudoroegneria|gb167|FF339936 539 79 90.3 blastp 294 LAB25_H3rye|gb164|BE493752 540 79 84.2 blastp 295 LAB25_H4 wheat|gb164|TAU73210541 79 89.2 blastp 296 LAB25_H5 wheat|gb164|TAU73211 542 79 91.5 blastp297 LAB25_H6 wheat|gb164|WHTWCOR 543 79 90.1 blastp 298 LAB31_H0b_oleracea|gb161|AM387244 544 81 94.6 blastp 299 LAB31_H1b_rapa|gb162|AT000569 545 81 97.9 blastp 300 LAB31_H2canola|gb161|DY012596 546 81 95.2 blastp 301 LAB31_H3radish|gb164|EV537620 547 81 87.1 blastp 302 LAB32_H0pseudoroegneria|gb167|FF342820 548 82 89.1 blastp 303 LAB33_H0sugarcane|gb157.3|BQ530200 549 83 88.1 blastp 304 LAB34_H0bean|gb167|BQ481761 550 84 86.1 blastp 305 LAB34_H1soybean|gb168|AI755294 551 84 90.9 blastp 306 LAB35_H0barley|gb157.3|Y07823 552 85 92.4 blastp 307 LAB35_H1brachypodium|gb169|Y07823 553 85 83.96 tblastn 308 LAB36_H0maize|gb170|DW878104 554 86 88.3 blastp 309 LAB38_H0barley|gb157.3|AL450676 555 87 91 blastp 310 LAB38_H1barley|gb157.3|BE438884 556 87 91.4 blastp 311 LAB38_H2barley|gb157.3|BF625343 557 87 84.9 blastp 312 LAB38_H3barley|gb157.3|BG299345 558 87 86.12 tblastn 313 LAB38_H4leymus|gb166|CD808961 559 87 94.3 blastp 314 LAB38_H5pseudoroegneria|gb167|FF340156 560 87 95.9 blastp 315 LAB38_H6wheat|gb164|AF495872 561 87 97.1 blastp 316 LAB38_H7wheat|gb164|BE591570 562 87 98.4 blastp 317 LAB39_H0switchgrass|gb167|FE640133 563 88 84.2 blastp 318 LAB39_H1switchgrass|gb167|FL746283 564 88 84.6 blastp 319 LAB39_H2wheat|gb164|CA484141 565 88 99.2 blastp 320 LAB40_H0barley|gb157.3|BF258976 566 89 96.9 blastp 321 LAB40_H1brachypodium|gb169|BE488436 567 89 84.4 blastp 322 LAB41_H0barley|gb157.3|BE437787 568 90 90.5 blastp 323 LAB43_H0leymus|gb166|EG377283 569 91 92.5 blastp 324 LAB43_H1wheat|gb164|AL822945 570 91 91.7 blastp 325 LAB43_H2wheat|gb164|BE413988 571 91 93.2 blastp 326 LAB43_H3wheat|gb164|CA610190 572 91 92.8 blastp 327 LAB45_H0apple|gb171|CN488819 573 92 80 blastp 328 LAB45_H1barley|gb157.3|AL502429 574 92 88.8 blastp 329 LAB45_H2basilicum|gb157.3|DY328093 575 92 80.7 blastp 330 LAB45_H3brachypodium|gb169|BE403542 576 92 90.3 blastp 331 LAB45_H4cotton|gb164|AI727046 577 92 80.42 tblastn 332 LAB45_H5fescue|gb161|DT697400 578 92 83.1 blastp 333 LAB45_H6leymus|gb166|EG380210 579 92 87.9 blastp 334 LAB45_H7maize|gb170|LLAI855293 580 92 96.4 blastp 335 LAB45_H8medicago|09v1|AW690268 581 92 80.76 tblastn 336 LAB45_H9poplar|gb170|BI124748 582 92 80.5 blastp 337 LAB45_H10potato|gb157.2|BF053337 583 92 80 tblastn 338 LAB45_H11rice|gb170|OS02G50350 584 92 92.4 blastp 339 LAB45_H12switchgrass|gb167|FL865538 585 92 96.2 blastp 340 LAB45_H13tomato|gb164|BG126074 586 92 80.19 tblastn 341 LAB45_H14wheat|gb164|BE403542 587 92 88.8 blastp 342 LAB51_H0barley|gb157.3|BE421767 588 95 85.2 blastp 343 LAB51_H1wheat|gb164|CA615952 589 95 93.1 blastp 344 BDL103_H0barley|gb157.3|BI954496 590 96 82.75 tblastn 345 BDL103_H1barley|gb157.3|BI956043 591 96 83.53 tblastn 346 BDL103_H2brachypodium|gb169|BE497565 592 96 85.1 blastp 347 BDL103_H3leymus|gb166|EG378510 593 96 84.6 blastp 348 BDL103_H4pseudoroegneria|gb167|FF346555 594 96 81.89 tblastn 349 BDL103_H5wheat|gb164|BE497565 595 96 82.68 tblastn 350 BDL103_H6wheat|gb164|BF428885 596 96 83.14 tblastn 351 BDL166_H0b_rapa|gb162|CX267860 597 100 81.77 tblastn 352 BDL166_H1canola|gb161|CD820129 598 100 90.4 blastp 353 CTF113_H0castorbean|09v1|XM002524611 599 103 81 blastp 354 CTF113_H1poplar|gb170|BI124993 600 103 80.6 blastp 355 CTF113_H2poplar|gb170|CV228068 601 103 81.1 blastp 356 CTF180_H0castorbean|09v1|EG657203 602 106 80.7 blastp 357 CTF215_H0castorbean|09v1|XM002514996 603 108 81.2 blastp 358 CTF215_H1poplar|gb170|AI162434 604 108 82.3 blastp Table 6: Provided arepolynucleotides and polypeptides which are homologous to the identifiedpolynucleotides or polypeptides of Table 5. Homol. = homologue; Algor. =Algorithm; Polynucl. = polynucleotide; Polypep. = polypeptide. Homologywas calculated as % of identity over the aligned sequences. The querysequences were polynucleotide sequences SEQ ID NOs: 1-59 and 638) orpolypeptides sequences SEQ ID NOs: 60-112, and the subject sequences areprotein sequences identified in the database based on greater than 80%identity to the predicted translated sequences of the query nucleotidesequences.

Example 3 Gene Cloning and Generation of Binary Vectors for PlantExpression

To validate their role in improving ABST, yield, growth rate, vigor,biomass, nitrogen use efficiency and/or oil content selected genes wereover-expressed in plants, as follows.

Cloning Strategy

Genes listed in Examples 1 and 2 hereinabove were cloned into binaryvectors for the generation of transgenic plants. For cloning, thefull-length open reading frames (ORFs) were identified. EST clusters andin some cases mRNA sequences were analyzed to identify the entire openreading frame by comparing the results of several translation algorithmsto known proteins from other plant species.

In order to clone the full-length cDNAs, reverse transcription (RT)followed by polymerase chain reaction (PCR; RT-PCR) was performed ontotal RNA extracted from leaves, roots or other plant tissues, growingunder normal conditions. Total RNA extraction, production of cDNA andPCR amplification was performed using standard protocols describedelsewhere (Sambrook J., E. F. Fritsch, and T. Maniatis. 1989. MolecularCloning. A Laboratory Manual, 2nd Ed. Cold Spring Harbor LaboratoryPress, New York.) which are well known to those skilled in the art. PCRproducts were purified using PCR purification kit (Qiagen)

Usually, 2 sets of primers were prepared for the amplification of eachgene, via nested PCR (meaning first amplifying the gene using externalprimers and then using the produced PCR product as a template for asecond PCR reaction, where the internal set of primers are used).Alternatively, one or two of the internal primers were used for geneamplification, both in the first and the second PCR reactions (meaningonly 2-3 primers were designed for a gene). To facilitate furthercloning of the cDNAs, an 8-12 bp extension was added to the 5′ of eachinternal primer. The primer extension includes an endonucleaserestriction site. The restriction sites were selected using twoparameters: (a) the restriction site does not exist in the cDNAsequence; and (b) the restriction sites in the forward and reverseprimers are designed such that the digested cDNA is inserted in thesense direction into the binary vector utilized for transformation. InTable 7 below, primers used for cloning selected genes are provided.

TABLE 7 PCR primers for cloning selected genes of the inventionRestriction Gene Enzymes used  Primers used for Name for cloningamplification (SEQ ID NOs:) BDL103S SalI, XbaIBDL103_Short_F_SalI SEQ ID NO: 677 AATGTCGACTCTGGGCTCAGGGATAGGBDL103_NR_XbaI SEQ ID NO: 678 TATCTAGACTACTAAAAGGAATTATCTAGCAGAGG BDL12SalI, SacI BDL12_gDNA_NF_SalI SEQ ID NO: 679AATGTCGACGTTCTATCCCCAACTCTAAATG BDL12_gDNA_NR_SacI SEQ ID NO: 680AGAGCTCCTTAAAGTTCTATCGAGATAGTGC BDL14 SalI, XbaIBDL14_ORF_F1_SalI SEQ ID NO: 681 AATGTCGACAACAATGGATCTACAACAGTCCGAAACBDL14_ORF_F1_SalI SEQ ID NO: 681 AATGTCGACAACAATGGATCTACAACAGTCCGAAACBDL14_ORF_NR_XbaI SEQ ID NO: 682 AATCTAGACACTCAGACAGCTGGGTATTAAACBDL14_ORF_ER_SacI SEQ ID NO: 683 AGAGCTCGTTGTGGCACTCAGACAGCTG BDL166XbaI, SacI BDL166_NF_XbaI SEQ ID NO: 684AATCTAGAAAAGTTACACCTTACTAAACACAAAC BDL166_NR_SacI SEQ ID NO: 685TGAGCTCTCTTGTTGATAGTCTTCATAATCG BDL210 SalI, XbaIBDL210_NF_SalI SEQ ID NO: 686 AAAGTCGACAACAAAGTTATGGGTTTCTCGBDL210_EF_SalI SEQ ID NO: 687 AAAGTCGACGAGCAACAAAGTTATGGGTTTCBDL210_NR_XbaI SEQ ID NO: 688 ATTCTAGATTAGGATGATCAGGAGATGAGAGAGBDL210_ER_XbaI SEQ ID NO: 689 ATTCTAGACTAAAGTAGAGAGATGGATGATCAGG CTF113CTF113_ORF_F_Sm SEQ ID NO: 690 GACCCGGGAAACGATGGAGGATCTTGCCCTF113_ORF_R_Sc SEQ ID NO: 691 CAGAGCTCTTGGAATTGAAATGTCATTACAGAG CTF163SalI, XbaI CTF163_NF_SalI SEQ ID NO: 692 AAAGTCGACGAACTGGTTGTTCTTGGCTATGCTF163_NR_XbaI SEQ ID NO: 693 ATTCTAGACCAGATGAACTTGGCTTTATC CTF175EcoRV, SacI CTF175_ORF_NF_EcRV SEQ ID NO: 694AGGATATCTTTCGATCACCGTGATGGC CTF175_ORF_EF_EcRV SEQ ID NO: 695AAGATATCAGAGCATTTCGATCACCGTG CTF175_ORF_NR_Sc SEQ ID NO: 696GCGAGCTCGTAGTGACGTCACCGGTTC CTF175_ORF_ER_Sc SEQ ID NO: 697TCGAGCTCCTCACCTTTCACTATCACCC CTF180 SalI, SacICTF180_NF_SalI SEQ ID NO: 698 AAAGTCGACTTCGATGTGGGATAACTGAATCCTF180_ER_SacI SEQ ID NO: 699 AACGAGCTCATTCAACAACCTAACCATCTTTGCTF180_NR_SacI SEQ ID NO: 700 AATGAGCTCTTTTCTTTACAGTGGAATCTGCCTF180_ER_SacI SEQ ID NO: 699 AACGAGCTCATTCAACAACCTAACCATCTTTG CTF205CTF205_EF_SalI SEQ ID NO: 701 AAAGTCGACGAAAACACAGATGGAAGATATTAAACCTF205_ER_XbaI SEQ ID NO: 702 ATTCTAGATGGACTTACAGGTCAAGAAGGTAG CTF215SalI, XbaI CTF215_NF_SalI SEQ ID NO: 703 AAAGTCGACAAGTTTGGAAAGAGATGAATCCCTF215_NR_XbaI SEQ ID NO: 704 ATTCTAGACTAAGCAAGCAGAAACAAAATATAGC CTF226SalI, XbaI CTF226_NF_SalI SEQ ID NO: 705 AAAGTCGACGCCAAGGTCAAACGAAGGCTF226_EF_SalI SEQ ID NO: 706 AAAGTCGACCAAAAGCCAAGGTCAAACGCTF226_NR_XbaI SEQ ID NO: 707 ATTCTAGACTAAACTTATGCAACATGAGCTGGCTF226_ER_XbaI SEQ ID NO: 708 ACTCTAGAAAGTCATTATCCTAGTTCAGTTTGC LAB11SalI, XbaI LAB11_NF_SalI SEQ ID NO: 709 AAAGTCGACATCTACTGCCTTTGACCGATGLAB11_NR_XbaI SEQ ID NO: 710 AATTCTAGATTACAGTTAAGTGAGGACATTCTTGG LAB13SalI, XbaI LAB13_NF_SalI SEQ ID NO: 711 AAAGTCGACCCCAAGATCGATATAAATTTCCLAB13_NR_XbaI SEQ ID NO: 712 AACTCTAGAAACCACCATGCTTGCTCATC LAB14EcoRV, EcoRV LAB14_NF_EcoRV SEQ ID NO: 713 AATGATATCTTCCATTGTTACACGCGTTCLAB14_NR_EcoRV SEQ ID NO: 714 AATGATATCTTAGGTGATTTAAAGCCAGAGGG LAB16SalI, XbaI LAB16_NF_SalI SEQ ID NO: 715 AAAGTCGACAACCAGACAAGAGAGAAAACAAGLAB16_NR_XbaI SEQ ID NO: 716 AATTCTAGATTACAATCACATAACAGAACAAGCAG LAB17EcoRV, PstI LAB17_NF_EcoRV SEQ ID NO: 717 AATGATATCTTGTTTCGTTTTCCCTTAGCLAB17_NR_PstI SEQ ID NO: 718 AATCTGCAGTCACCAGTTCACCACCATCTAC LAB2EcoRV, PstI LAB2_NF_EcoRV SEQ ID NO: 719 AATGATATCTTGCCGGTCGATCTTGAGLAB2_EF_EcoRV SEQ ID NO: 720 AATGATATCCCTATATCTCCCTCCTCCTCC LAB2LAB2_NR_PstI SEQ ID NO: 721 AATCTGCAGTCAGCCACGGACTACCTACATGACLAB2_ER_PstI SEQ ID NO: 722 AACCTGCAGACAATTTCATTCTGTGGGTTC LAB20 SmaILAB2O_NF SEQ ID NO: 723  CCTCAGAAAATCACCGTACGLAB2O_NR_SmaI SEQ ID NO: 724 TAACCCGGGCCTATGAACAGATATCTGACATGATC LAB21SalI, XbaI LAB21_NF_SalI SEQ ID NO: 725TTAGTCGACGGAGAGAGATCTTCTAGCTACATAC LAB21_NR_XbaI SEQ ID NO: 726TAATCTAGATCACAGGACAGGACACCATCAAC LAB22 SalI, XbaILAB22_NF_SalI SEQ ID NO: 727 TTAGTCGACGGAGACAAAGATGGAGAACAACLAB22_NR_XbaI SEQ ID NO: 728 TATTCTAGACCGAAATTAAACAACAAGTACAC LAB23EcoRV, EcoRV LAB23_NF_EcoRV SEQ ID NO: 729AAAGATATCGGAGGTACATATAGCTAGCGAAG LAB23_NR_EcoRV SEQ ID NO: 730AATGATATCCTAACAAAATCCACGACTCCACTG LAB24 SalI, XbaILAB24_NF_SalI SEQ ID NO: 731 AAAGTCGACGAGAGAGGATGGTGAGCAGCLAB24_NR_XbaI SEQ ID NO: 732 AATTCTAGATTACGTGTAGTCATCAAATCACGC LAB25SalI, XbaI LAB25_NF_SalI SEQ ID NO: 733 AATGTCGACTCTAGCTCCCACGAGTCTTTAGLAB25_NR_XbaI SEQ ID NO: 734 AATTCTAGATTACAACAATTTAATGGAGGTCCG LAB3SalI, XbaI LAB3_NF_SalI SEQ ID NO: 735 TTAGTCGACGAGCAAAAAATGAAGGAGAACLAB3_NR_XbaI SEQ ID NO: 736 TATTCTAGATTACAGAGATTGTTAAGGTTGGACC LAB31EcoRV, PstI LAB31 NF EcoRV SEQ ID NO: 737AAAGATATCTCACAATTTCATTCACAAGTCG LAB31 NR PstI SEQ ID NO: 738AATCTGCAGTTTTCAAATCCAAACCCAAC LAB32 SalI, XbaILAB32_NF_SalI SEQ ID NO: 739 AAAGTCGACCTTTCCTTTCCTTTCCATCCLAB32_EF_SalI SEQ ID NO: 740 AATTCTAGAAGCCATCACCACGCATTACLAB32_NR_XbaI SEQ ID NO: 741 AATTCTAGAAGCACTGAGCAGCCTTCATCLAB32_ER_XbaI SEQ ID NO: 740 AATTCTAGAAGCCATCACCACGCATTAC LAB33EcoRV, EcoRV LAB33 NF EcoRV SEQ ID NO: 742 TCAGATATCCATCGCATCGCATCCATCLAB33 NR EcoRV SEQ ID NO: 743 ATAGATATCGCTGCCTGCTTCTGATCTG LAB34SalI, XbaI LAB34_NF_SalI SEQ ID NO: 744 AAAGTCGACGCTAGTGAGATACCATGGACAACLAB34_NR_XbaI SEQ ID NO: 745 AAATCTAGATTACTTCTATGCTGGAATGACTTTG LAB35LAB35_NF_SalI SEQ ID NO: 746 AAAGTCGACCAGATCGCGATGAAGTCTTGLAB35_EF_SalI SEQ ID NO: 747 AAAGTCGACAGGGGAGAAGAGAGAGAGACAGLAB35_NR_XbaI SEQ ID NO: 748 AAATCTAGATTAGCTCGTTCATTTAGCCTCAGLAB35 ER Xba SEQ ID NO: 749 TCCTCTAGAGAGTTTATTCCTCGACGATGC LAB36LAB36_NF_SalI SEQ ID NO: 750 AAAGTCGACCAGTGTAGAGCAAGAGGTGTGGLAB36_EF_SalI SEQ ID NO: 751 AAAGTCGACTCGTCTCGATCAGTGTAGAGCLAB36_NR_XbaI SEQ ID NO: 752 AAATCTAGATTACGTCGTTCATTTAGCCTTTGLAB36_ER_XbaI SEQ ID NO: 753 AATTCTAGACAATTATTCCACAGGACATCAC LAB38 EcoRVLAB38_NF_EcoRV SEQ ID NO: 754 AAAGATATCAGGAGATATGGCCCAGAGGLAB38_EF_EcoRV SEQ ID NO: 755 TTAGATATCCTGCTTGCAATACTTAGTAGAGGLAB38_NR_EcoRV SEQ ID NO: 756 AAAGATATCTTAACGTACTCTCAGGTGAGGCGLAB38_ER_EcoRV SEQ ID NO: 757 TAAGATATCTTTATTTATTCACCGGAGCAAC LAB39SalI, XbaI LAB39_NF_SalI SEQ ID NO: 758 AAAGTCGACCAAAATAGCAGAGATGGGAGGLAB39_NR_XbaI SEQ ID NO: 759 AAATCTAGATCACGGTAATCAGTTCAGCATGG LAB40SalI, XbaI LAB40_NF_SalI SEQ ID NO: 760 AAAGTCGACACACTACCAACATGGAAACATACLAB40_EF_SalI SEQ ID NO: 761 AAAGTCGACGCTGAATCGGCACACACTACLAB40_NR_XbaI SEQ ID NO: 762 AATTCTAGATGACCATCATCAGTTCATTGCLAB40_ER_XbaI SEQ ID NO: 763 AATTCTAGAGGAGTGAGGACTTTACAAAATG LAB41SalI, XbaI LAB41_NF_SalI SEQ ID NO: 764 AAAGTCGACAAGAGCTGCGAGAGGAAGGLAB41_NR_XbaI SEQ ID NO: 765 AAATCTAGATTAACATCAATTGTCAGTCATCGG LAB45SalI, StuI LAB45_NF_SalI SEQ ID NO: 766 AAAGTCGACATTCTTATCAAAACAGAGGAACCLAB45_EF_SalI SEQ ID NO: 767 AAAGTCGACCTCCCTCAGATTCTTATCAAAACLAB45_NR_XbaI SEQ ID NO: 768 AAATCTAGATTAGCATCAGTTGGATACCATGLAB45_ER_XbaI SEQ ID NO: 769 AAATCTAGATTAAGTCACAAGTTGAAGCATGGTG LAB49EcoRV, EcoRV LAB49_NF_EcoRV SEQ ID NO: 770 AAAGATATCACGATCAGCCATGAAGAGCLAB49_NR_EcoRV SEQ ID NO: 771 AAAGATATCTTATTAAGCTGGCTGGTTGTGAC LABSLAB5_EF_EcoRV SEQ ID NO: 772 AAAGATATCCTCTTCCACAATCCACATTCCLAB5_ER_PstI SEQ ID NO: 773 AATCTGCAGTGACGATCCATCTATGAACAAC LAB50LAB5O_NF_SalI SEQ ID NO: 774 AAAGTCGACCACGGAGAAAAGAAAGATCGLAB5O_NR_XbaI SEQ ID NO: 775 AAATCTAGATTAAAACTCCGGCTGCTAGACC LAB51SalI, XbaI LAB51_NF_SalI SEQ ID NO: 776 AAAGTCGACAGTACTTCGGTTGATGGCTTCLAB51_EF_SalI SEQ ID NO: 777 AAAGTCGACCTCTGCTCGTCTCTGCATTTAG LAB51LAB51_NR_XbaI SEQ ID NO: 778 AAATCTAGATTAAACACTTATGTATGCACGCTTAGLAB51_ER_XbaI SEQ ID NO: 779 AAATCTAGATTATCCACACCAAGACCAAGACAG Providedare primers and the restriction sites and enzymes used for cloningselected genes (polynucleotides, provided by gene name) identifiedherein.

TABLE 8 Restriction enzymes and cloning vectors used to clone selectedgenes of the invention Restriction enzymes Restriction enzymes used forcloning into used for cloning into Restriction enzymes Gene binaryvector- binary vector- used for digesting name Binary vector FORWARDREVERSE the binary vector BDL103 pBXYN (pGI_35S) XbaI Sac I XbaI, Sac IBDL103 pBXYN (pGI_35S) SalI EcoR I SalI, EcoR I BDL11 pM (pMBLArt) NotINot I NotI, Not I BDL12 pBXYN (pGI_35S) HindIII EcoR I HindIII, EcoR IBDL14 pBXYN (pGI_35S) HindIII EcoRI HindIII, EcoRI BDL166 pQXYN XbaIEcoRI XbaI, EcoRI BDL17 pM (pMBLArt) NotI Not I NotI, Not I BDL17S pM(pMBLArt) NotI Not I NotI, Not I BDL210 pQXYN SalI EcoRI SalI, EcoRICTF113 pBXYN (pGI_35S) SmaI Sac I SmaI, SacI CTF163 pQXYN SalI SacISalI, SacI CTF175 pBXYN (pGI_35S) EcoRV Sac I SmaI, SacI CTF180 pQXYNSalI EcoRI SalI, EcoRI CTF205 pQXYN SalI SacI SalI, SacI CTF215 pQXYNSalI SacI SalI, SacI CTF226 pQXYN SalI SacI SalI, SacI LAB11 pQFN SalIEcoRI SalI, EcoRI LAB13 pQFN SalI EcoRI SalI, EcoRI LAB14 pQYN BamHISmaI BamHI, Ecl136II LAB15 pQFN SalI Ecl136II SalI, StuI LAB16 pQFN SalIEcoRI SalI, EcoRI LAB17 pQFN EcoRV SmaI StuI, StuI LAB18 pQFN SalIEcl136II SalI, StuI LAB2 pQFN EcoRV SmaI StuI, StuI LAB20 pQYN HindIIISmaI HindIII, Ecl136II LAB21 pQFN SalI EcoRI SalI, EcoRI LAB22 pQFN SalIXbaI SalI, XbaI LAB23 pQFN EcoRV EcoRV StuI, StuI LAB24 pQYN_6669 SalIEcoRI SalI, EcoRI LAB25 pQFN SalI EcoRI SalI, EcoRI LAB3 pQFN SalI EcoRISalI, EcoRI LAB31 pQYN BamHI SmaI BamHI, Ecl136II LAB32 pQFN SalI EcoRISalI, EcoRI LAB33 pQFN EcoRV EcoRV StuI, StuI LAB34 pQFN SalI EcoRISalI, EcoRI LAB35 pQFN SalI Ecl136II SalI, StuI LAB36 pQFN SalI EcoRVSalI, StuI LAB38 pQYN BamHI SmaI BamHI, Ecl136II LAB39 pQFN SalI EcoRISalI, EcoRI LAB4 pQFN EcoRV EcoRV SmaI, SmaI LAB40 pQFN SalI EcoRI SalI,EcoRI LAB41 pQYN_6669 SalI EcoRI SalI, EcoRI LAB45 pQFN SalI BamHI SalI,BamHI LAB49 pQFN EcoRV Ecl136II StuI, StuI LAB5 pQFN EcoRV KpnI StuI,KpnI LAB50 pQFN SalI BamHI SalI, BamHI LAB51 pQYN_6669 SalI EcoRI SalI,EcoRI LAB8 pQFN BamHI XhoI BamHI, XhoI LAB9 pQFN BamHI KpnI BamHI, KpnITable 8: Provided are the restriction enzymes and cloning vectors usedfor cloning selected genes of the invention.

TABLE 9 Primers used for colony screening of the binary plasmid ColonyScreening FP FP SEQ Colony Screening RP RP SEQ Gene Name Name ID NO:Name ID NO: BDL103_Long 35S_1F 780 NOS R 784 BDL103_ShortBDL103_Short_F_SalI 677 101_ER 785 BDL11_GA 35S_1F 780 101_R 786 BDL12101 EF 781 BDL12_gDNA_NR_SacI 680 BDL14 BDL14_ORF_F1_SalI 681 101_R 786BDL166 35S_1F 780 BDL166_NR_SacI 685 BDL17 35S_1F 780 101_R 786 BDL17101-F 782 BDL17_GA_R 787 BDL210 35S_1F 780 BDL210_NR_XbaI 688 CTF11335S_1F 780 NOS R 784 CTF163 35S_1F 780 CTF163_NR_XbaI 693 CTF175 35S_1F780 NOS R 784 CTF180 35S_1F 780 CTF180_NR_SacI 700 CTF205 35S_1F 780CTF205_ER_XbaI 702 CTF215 35S_1F 780 CTF215_NR_XbaI 704 CTF226 35S_1F780 CTF226_NR_XbaI 707 LAB11 6669 F 783 LAB11_NR_XbaI 710 LAB13 6669 F783 101_R 786 LAB14 p6669-F 783 101_ER 785 LAB15 p6669-F 783 101_ER 785LAB16 6669 F 783 LAB16_NR_XbaI 716 LAB17 p6669-F 783 LAB17_NR_PstI 718LAB18 p6669-F 783 101_ER 785 LAB2 p6669-F 783 LAB2_NR_PstI 721 LAB20101-F 782 LAB20_NR_SmaI 724 LAB21 6669 F 783 LAB21_NR_XbaI 726 LAB226669 F 783 LAB22_NR_XbaI 728 LAB23 p6669-F 783 LAB14_NR_EcoRV 714 LAB246669 F 783 LAB24_NR_XbaI 732 LAB25 6669 F 783 LAB25_NR_XbaI 734 LAB36669 F 783 LAB3_NR_XbaI 736 LAB31 LAB31 NF EcoRV 737 101 EF 781 LAB326669 F 783 LAB32_NR_XbaI 741 LAB33 p6669-F 783 LAB33_R1_seq 788 LAB346669 F 783 LAB34_NR_XbaI 745 LAB35 6669 F 783 LAB35_NR_XbaI 748 LAB366669 F 783 LAB36_NR_XbaI 752 LAB38 LAB38_NF_EcoRV 754 101 EF 781 LAB396669 F 783 LAB39_NR_XbaI 759 LAB4 6669 F 783 LAB4_R_GA 789 LAB40 6669 F783 LAB40_NR_XbaI 762 LAB41 6669 F 783 LAB41_NR_XbaI 765 LAB45 6669 F783 LAB45_NR_XbaI 790 LAB49 6669 F 783 LAB49_NR_EcoRV 771 LAB5 p6669-F783 101_ER 785 LAB50 6669 F 783 LAB50_NR_XbaI 775 LAB51 6669 F 783LAB51_NR_XbaI 778 LAB8 6669 F 783 LAB8_GA_rev 791 LAB9 6669 F 783LAB9_GA_rev 792 Table 9. Provided are the forward primers (FP) andreverse primers (RP) along with their sequence identifiers used forscreening of colonies harboring the cloned genes of some embodiments ofthe invention.

TABLE 10 Cloned genes from cDNA libraries or genomic DNA and thepolypeptides encoded thereby Polynuc. Polypep. Gene High copy Amplifiedfrom SEQ ID SEQ ID Name plasmid Organism Origin NO: NO: BDL103_LongGeneArt 670 96 BDL103_Short pGXN RICE Oryza sativa cDNA- 671 672 (pKG +Nos + 35S) L. Japonica ND RICE BDL11 pGXN GeneArt 639 661 (pKG + Nos +35S) BDL12 pGXN ARABIDOPSIS gDNA 640 662 (pKG + Nos + 35S) Arabidopsisthaliana ND BDL14 pGXN ARABIDOPSIS cDNA 641 99 (pKG + Nos + 35S)Arabidopsis thaliana ND BDL166 pGXN ARABIDOPSIS cDNA 642 100 (pKG +Nos + 35S) Arabidopsis thaliana ND BDL17 pGXN GeneArt 643 101 (pKG +Nos + 35S) BDL17 pGN_Napin GeneArt 643 101 BDL210 pGXN ARABIDOPSIS cDNA644 102 (pKG + Nos + 35S) Arabidopsis thaliana ND CTF113 pKS(Pks_J)Cotton cDNA 645 663 CTF163 pGXN COTTON cDNA 646 664 (pKG + Nos + 35S)Gossypium barbadense ND CTF175 pKS(Pks_J) Cotton cDNA 647 665 CTF180pGXN COTTON cDNA 648 666 (pKG + Nos + 35S) Gossypium barbadense NDCTF205 pGXN COTTON cDNA 649 667 (pKG + Nos + 35S) Gossypium barbadenseND CTF215 pGXN COTTON cDNA 650 668 (pKG + Nos + 35S) Gossypiumbarbadense ND CTF226 pGXN COTTON cDNA 651 669 (pKG + Nos + 35S)Gossypium barbadense ND LAB11 pGXN RICE Oryza sativa cDNA 609 65 (pKG +Nos + 35S) L. Japonica ND LAB13 pGXN RICE Oryza sativa cDNA 610 66(pKG + Nos + 35S) L. Japonica ND LAB14 pKSJ_6669a RICE Oryza sativa cDNA611 67 L. Japonica ND LAB15 Gene Art 612 68 LAB16 pGXN COTTON cDNA 61470 (pKG + Nos + 35S) Gossypium barbadense ND LAB17 pKSJ_6669a SORGHUMcDNA 615 71 Sorghum bicolor Monsanto S5 LAB18 GeneArt 616 72 LAB2pKS(Pks_J) BARLEY cDNA 613 69 Hordeum vulgare L. ND LAB20 pUC19_pr6669RICE Oryza sativa cDNA 617 73 L. Japonica ND LAB21 pGXN BARLEY cDNA 618653 (pKG + Nos + 35S) Hordeum vulgare L. ND LAB22 pGXN SORGHUM cDNA 61975 (pKG + Nos + 35S) Sorghum bicolor Monsanto S5 LAB23 pKSJ_6669a BARLEYcDNA 621 77 Hordeum vulgare L. ND LAB24 pGXN SORGHUM cDNA 622 655 (pKG +Nos + 35S) Sorghum bicolor Monsanto S5 LAB25 pGXN BARLEY cDNA 623 656(pKG + Nos + 35S) Hordeum vulgare L. ND LAB3 pGXN COTTON cDNA 620 654(pKG + Nos + 35S) Gossypium hirsutum Akala LAB31 pKSJ_6669a COTTON cDNA624 81 Gossypium hirsutum Akala LAB32 pGXN BARLEY cDNA 625 82 (pKG +Nos + 35S) Hordeum vulgare L. ND LAB33 pKS(Pks_J) SORGHUM cDNA 626 83Sorghum bicolor Monsanto S5 LAB34 pGXN SOYBEAN cDNA 627 657 (pKG + Nos +35S) Glycine max ND LAB35 Topo B WHEAT cDNA 628 658 Triticum aestivum L.ND LAB36 Topo B SORGHUM cDNA 629 86 Sorghum bicolor Monsanto S5 LAB38pKSJ_6669a WHEAT cDNA 630 87 Triticum aestivum L. ND LAB39 pGXN SORGHUMcDNA 631 659 (pKG + Nos + 35S) Sorghum bicolor Monsanto S5 LAB4 GeneArt605 60 LAB40 pGXN SORGHUM cDNA 632 660 (pKG + Nos + 35S) Sorghum bicolorMonsanto S5 LAB41 pGXN WHEAT cDNA 633 90 (pKG + Nos + 35S) Triticumaestivum L. ND LAB45 TopoB_LAB45 SORGHUM cDNA + part 634 92 Sorghumbicolor from GA Monsanto S5 LAB49 pKSJ_6669a RICE Oryza sativa cDNA 63593 L. Japonica ND LAB5 Topo B SORGHUM cDNA 606 652 Sorghum bicolorMonsanto S5 LAB50 Topo B RICE Oryza sativa cDNA 636 94 L. Japonica NDLAB51 pGXN WHEAT cDNA 637 95 (pKG + Nos + 35S) Triticum aestivum L. NDLAB8 GeneArt 607 63 LAB9 GeneArt 608 64 Table 10. Provided are thecloned and synthetic genes, the polypeptides encoded thereby along withtheir sequence identifiers. Also provided are the source of DNA used forcloning (cDNA or genomic DNA) and the organism from which the gene wascloned. Polynuc. = polynucleotide; Polypep. = polypeptide. BDL103 shortwas amplified from pGXN_BDL103. pGXN_BDL103 was amplified from cDNA-RICE Oryza sativa L. Japonica ND. LAB45 was composed of a part clonedfrom cDNA with the primers indicated and a part ordered from GA

PCR products were digested with the restriction endonucleases (Roche,Switzerland) according to the sites design in the primers (Table 7).Each digested PCR product was inserted into a high copy vectororiginated from pBlue-script KS plasmid vector (pBlue-script KS plasmidvector, Hypertext Transfer Protocol://World Wide Web (dot) stratagene(dot) com/manuals/212205 (dot) pdf) or pUC19 (New England BioLabs Inc).In case of the high copy vector originated from pBlue-script KS plasmidvector (pGXN) the PCR product was inserted in the high copy plasmidupstream to the NOS terminator (SEQ ID NO:673) originated from pBI 101.3binary vector (GenBank Accession No. U12640, nucleotides 4417 to 4693)and down stream to the 35S promoter (SEQ ID NO:675). In other cases(pKSJ_(—)6669a or pUC19_pr6669) the At6669 promoter (SEQ ID NO:674) wasalready cloned into the pBlue-script KS or pUC19 respectively, so thegene was introduced downstream of the promoter.

Sequencing of the inserted genes was performed, using the ABI 377sequencer (Applied Biosystems). In all the cases, after confirming thesequences of the cloned genes, the cloned cDNA accompanied with the NOSterminator was introduced into a modified pGI binary vector containingthe At6669 promoter via digestion with appropriate restrictionendonucleases. In other cases the cloned cDNA accompanied with theAt6669 promoter was introduced into a pGI vector (that hasn't alreadycontained the At6669 promoter). In any case the insert was followed bysingle copy of the NOS terminator (SEQ ID NO: 673). Part of the geneswere introduced into a binary vector pGI containing the 35S promoter.The digested products and the linearized plasmid vector were ligatedusing T4 DNA ligase enzyme (Roche, Switzerland).

Several DNA sequences of the selected genes were synthesized by GeneArt(Hypertext Transfer Protocol://World Wide Web (dot) geneart (dot) com/).Synthetic DNA is designed in silico. Suitable restriction enzymes sitesare added to the cloned sequences at the 5′ end and at the 3′ end toenable later cloning into the desired binary vector.

The pPI plasmid vector is constructed by inserting a synthetic poly-(A)signal sequence, originating from pGL3 basic plasmid vector (Promega,GenBank Accession No. U47295; nucleotides 4658-4811) into the HindIIIrestriction site of the binary vector pBI101.3 (Clontech, GenBankAccession No. U12640). pGI (FIG. 1) is similar to pPI, but the originalgene in the back bone is GUS-Intron, rather than GUS.

The modified pGI vector (FIG. 2) is a modified version of the pGI vectorin which the cassette is inverted between the left and right borders sothe gene and its corresponding promoter are close to the right borderand the NPTII gene is close to the left border.

At6669, the Arabidopsis thaliana promoter sequence (set forth in SEQ IDNO: 674) was inserted in the pGI binary vector, upstream to the clonedgenes, followed by DNA ligation and binary plasmid extraction frompositive E. coli colonies, as described above. Colonies were analyzed byPCR using the primers covering the insert which were designed to spanthe introduced promoter and gene. Positive plasmids were identified,isolated and sequenced as described above.

Some genes were cloned downstream of the Napin promoter (SEQ ID NO:676)and upstream to the NOS terminator in the pMBLArt vector. The vectordisplays resistance to Basta.

Promoters used: Arabidopsis At6669 promoter (SEQ ID NO: 674; which isSEQ ID NO: 61 of WO04081173), Napin (SEQ ID NO: 676) and 35S (SEQ ID NO:675).

Example 4 Transforming Agrobacterium Tumefaciens Cells with BinaryVectors Harboring Putative Genes

Each of the binary vectors described in Example 3 above were used totransform Agrobacterium cells. Two additional binary constructs, havinga GUS/Luciferase reporter gene replacing the selected gene (positioneddownstream of the At6669 promoter), were used as negative controls.

The binary vectors were introduced to Agrobacterium tumefaciens GV301,or LB4404 competent cells (about 10⁹ cells/mL) by electroporation. Theelectroporation was performed using a MicroPulser electroporator(Biorad), 0.2 cm cuvettes (Biorad) and EC-2 electroporation program(Biorad). The treated cells were cultured in LB liquid medium at 28° C.for 3 hours, then plated over LB agar supplemented with gentamycin (50mg/L; for Agrobacterium strains GV301) or streptomycin (300 mg/L; forAgrobacterium strain LB4404) and kanamycin (50 mg/L) at 28° C. for 48hours. Agrobacterium colonies developed on the selective media wereanalyzed by PCR using the primers which are designed to span theinserted sequence in the pPI plasmid. The resulting PCR products wereisolated and sequenced as described in Example 3 above, to verify thatthe correct polynucleotide sequences were properly introduced to theAgrobacterium cells.

Example 5 Transformation of Arabidopsis Thaliana Plants with theIdentified Polynucleotides of the Invention

Arabidopsis thaliana Columbia plants (T0 plants) were transformedaccording to the Floral Dip procedure [Clough S J, Bent A F. (1998)Floral dip: a simplified method for Agrobacterium-mediatedtransformation of Arabidopsis thaliana. Plant J. 16(6): 735-43] andDesfeux C, Clough S J, Bent A F. (2000) [Female reproductive tissues arethe primary targets of Agrobacterium-mediated transformation by theArabidopsis floral-dip method. Plant Physiol. 123(3): 895-904] withminor modifications. Briefly, T₀ Plants were sown in 250 ml pots filledwith wet peat-based growth mix. The pots were covered with aluminum foiland a plastic dome, kept at 4° C. for 3-4 days, then uncovered andincubated in a growth chamber at 18-24° C. under 16/8 hour light/darkcycles. The T₀ plants were ready for transformation six days beforeanthesis.

Single colonies of Agrobacterium carrying the binary constructs weregenerated as described in Example 4 above. Colonies were cultured in LBmedium supplemented with kanamycin (50 mg/L) and gentamycin (50 mg/L).The cultures were incubated at 28° C. for 48 hours under vigorousshaking and then centrifuged at 4000 rpm for 5 minutes. The pelletscomprising the Agrobacterium cells were re-suspended in a transformationmedium containing half-strength (2.15 g/L) Murashige-Skoog (Duchefa);0.044 μM benzylamino purine (Sigma); 112 μg/L B5 Gambourg vitamins(Sigma); 5% sucrose; and 0.2 ml/L Silwet L-77 (OSI Specialists, CT) indouble-distilled water, at pH of 5.7.

Transformation of T₀ plants was performed by inverting each plant intoan Agrobacterium suspension, such that the above ground plant tissue wassubmerged for 3-5 seconds. Each inoculated T₀ plant was immediatelyplaced in a plastic tray, then covered with clear plastic dome tomaintain humidity and was kept in the dark at room temperature for 18hours, to facilitate infection and transformation. Transformed(transgenic) plants were then uncovered and transferred to a greenhousefor recovery and maturation. The transgenic T₀ plants were grown in thegreenhouse for 3-5 weeks until siliques were brown and dry. Seeds wereharvested from plants and kept at room temperature until sowing.

For generating T₁ and T₂ transgenic plants harboring the genes, seedscollected from transgenic T₀ plants were surface-sterilized by soakingin 70% ethanol for 1 minute, followed by soaking in 5% sodiumhypochloride and 0.05% triton for 5 minutes. The surface-sterilizedseeds were thoroughly washed in sterile distilled water then placed onculture plates containing half-strength Murashige-Skoog (Duchefa); 2%sucrose; 0.8% plant agar; 50 mM kanamycin; and 200 mM carbenicylin(Duchefa). The culture plates were incubated at 4° C. for 48 hours thentransferred to a growth room at 25° C. for an additional week ofincubation. Vital T₁ Arabidopsis plants were transferred to a freshculture plates for another week of incubation. Following incubation theT₁ plants were removed from culture plates and planted in growth mixcontained in 250 ml pots. The transgenic plants were allowed to grow ina greenhouse to maturity. Seeds harvested from T₁ plants were culturedand grown to maturity as T₂ plants under the same conditions as used forculturing and growing the T₁ plants.

Example 6 Transgenic Plants Overexpressing the Polynucleotides of SomeEmbodiments of the Invention Exhibit Increased ABST, NUE, Biomass and/orGrowth Rate

Plants transgenic to the polynucleotides of some embodiments of theinvention were assayed for fertilizer use efficiency in a tissue cultureassay.

Assay 1: Plant Growth at Nitrogen Deficiency Under Tissue CultureConditions—

The present inventors have found the nitrogen use efficiency (NUE) assayto be relevant for the evaluation of the ABST candidate genes, sincenitrogen limiting conditions encourage root elongation, increases rootcoverage and allows detecting the potential of the plant to generate abetter root system under drought conditions. In addition, there areindications in the literature that biological mechanisms of NUE anddrought tolerance are linked (Wesley et al., 2002 Journal of ExperimentBotany Vol 53, No. 366, pp. 13-25).

Surface sterilized seeds were sown in basal media [50% Murashige-Skoogmedium (MS) supplemented with 0.8% plant agar as solidifying agent] inthe presence of Kanamycin (for selecting only transgenic plants). Aftersowing, plates were transferred for 2-3 days for stratification at 4° C.and then grown at 25° C. under 12-hour light 12-hour dark daily cyclesfor 7 to 10 days. At this time point, seedlings randomly chosen werecarefully transferred to plates with nitrogen-limiting conditions: 0.5MS media in which the combined nitrogen concentration (NH₄NO₃ and KNO₃)is 0.75 mM (nitrogen deficient conditions) or 15 mM [Normal (optimal)nitrogen concentration]. Each plate contains 5 seedlings of same event,and 3-4 different plates (replicates) for each event. For eachpolynucleotide of the invention at least four independent transformationevents were analyzed from each construct. Plants expressing thepolynucleotides of the invention were compared to the averagemeasurement of the control plants (generated by transformation of plantwith an empty vector under the same promoter or a vector comprising theGUS reporter gene under the same promoter) used in the same experiment.

Digital Imaging—

A laboratory image acquisition system, which consists of a digitalreflex camera (Canon EOS 300D) attached with a 55 mm focal length lens(Canon EF-S series), mounted on a reproduction device (Kaiser RS), whichincluded 4 light units (4×150 Watts light bulb) and located in adarkroom, is used for capturing images of plantlets sawn in agar plates.

The image capturing process was repeated every 3-4 days starting at day1 till day 10 (see for example the images in FIGS. 3A-F).

An image analysis system was used, which consists of a personal desktopcomputer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ1.39 (Java based image processing program which was developed at theU.S. National Institutes of Health and freely available on the internetat Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Imageswere captured in resolution of 10 Mega Pixels (3888×2592 pixels) andstored in a low compression JPEG (Joint Photographic Experts Groupstandard) format. Next, analyzed data was saved to text files andprocessed using the JMP statistical analysis software (SAS institute).

Seedling Analysis—

Using the digital analysis seedling data was calculated, including leafarea, root coverage and root length.

The relative growth rate for the various seedling parameters wascalculated according to the following formulas V, VI and VII.

Relative growth rate of leaf area=Regression coefficient of leaf areaalong time course.  Formula V:

Relative growth rate of root coverage=Regression coefficient of rootcoverage along time course.  Formula VI:

Relative growth rate of root length=Regression coefficient of rootlength along time course.  Formula VII:

At the end of the experiment, plantlets were removed from the media andweighed for the determination of plant fresh weight. Plantlets were thendried for 24 hours at 60° C., and weighed again to measure plant dryweight for later statistical analysis. Growth rate was determined bycomparing the leaf area coverage, root coverage and root length, betweeneach couple of sequential photographs, and results were used to resolvethe effect of the gene introduced on plant vigor, under osmotic stress,as well as under normal or optimal conditions. Similarly, the effect ofthe gene introduced on biomass accumulation, under osmotic stress aswell as under optimal conditions, was determined by comparing theplants' fresh and dry weight to that of control plants (containing anempty vector or the GUS reporter gene under the same promoter). Fromevery construct created, 3-5 independent transformation events wereexamined in replicates.

Statistical Analyses—

To identify genes conferring significantly improved tolerance to abioticstresses or enlarged root architecture, the results obtained from thetransgenic plants were compared to those obtained from control plants.To identify outperforming genes and constructs, results from theindependent transformation events tested were analyzed separately. Toevaluate the effect of a gene event over a control the data was analyzedby Student's t-test and the p value was calculated. Results wereconsidered significant if p≦0.1. The JMP statistics software package wasused (Version 5.2.1, SAS Institute Inc., Cary, N.C., USA).

Experimental Results

The genes presented in Tables 11-18, hereinbelow, were found to increaseABST by improving root performance, plant growth characteristic andplant biomass when grown under limiting nitrogen growth conditions ascompared to control plants.

Tables 11-18 depict analyses of root growth (root length and rootcoverage; Tables 11 and 12); plant biomass (plant fresh, dry weight andleaf area; Tables 13 and 14); root growth rate (relative growth rate ofroot length and root coverage; Tables 15 and 16); and leaf area and leafarea growth rate (relative growth rate of leaf area; Tables 17 and 18)when grown under limiting nitrogen conditions [low nitrogen or nitrogendeficient conditions (0.75 mM N)] in plants overexpressing thepolynucleotides of some embodiments of the invention under theregulation of a constitutive promoter [35S (SEQ ID NO:675) or At6669(SEQ ID NO:674)]. Evaluation of each gene was performed by testing theperformance of several events. Some of the genes were evaluated in morethan one tissue culture assay and the second experiment confirmed thesignificant increment in plant biomass. Event with p-value <0.05 wasconsidered statistically significant.

TABLE 11 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant roots undernitrogen deficient conditions Gene Plant Root length [cm] Gene PlantRoot Coverage [cm²] Name Event # Ave. p-value % incr. Name Event # Ave.p-value % incr. BDL103 8033.1 6.07 2.24E−04 22.7 CTF113 5871.2 15.173.7E−02 19 BDL103 8033.4 5.55 1.28E−02 12.3 Control 12.79 0.00 Control4.95 BDL103 8033.1 8.63 1.3E−03 34 CTF163 11012.2 5.46 4.92E−02 16Control 6.45 0.00 CTF163 11012.7 5.97 3.81E−03 27 CTF163 11011.2 7.032.1E−02 42 Control 4.71 0.00 CTF163 11012.2 7.59 2.8E−02 53 CTF16311011.2 5.86 7.62E−03 36 CTF163 11012.7 8.03 5.5E−04 62 CTF163 11012.26.16 1.70E−03 43 Control 4.96 0.00 CTF163 11012.4 5.64 3.36E−02 31CTF226 10982.1 8.81 8.8E−03 63 CTF163 11012.7 6.52 6.74E−03 52 CTF22610982.3 10.72 9.1E−03 98 Control 4.30 Control 5.42 0.00 CTF226 10982.37.02 9.00E−06 38 CTF205 11972.3 6.81 7.3E−04 44 Control 5.08 0.00Control 4.74 0.00 CTF205 11972.3 5.49 4.87E−02 14 Control 4.83 0.00Table 11: Analyses of plant roots (root length and root coverage) oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35S;SEQ ID NO: 675) when grown under limiting nitrogen conditions [lownitrogen or nitrogen deficient conditions (0.75 mM N)] as compared tocontrol plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

TABLE 12 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant roots undernitrogen deficient conditions Gene Plant Root length [cm] Gene PlantRoot Coverage [cm²] Name Event # Ave. p-value incr. Name Event # Ave.p-value % incr. LAB31 11423.4 6.72 5.38E−03 17 LAB31 11423.4 9.253.9E−03 47 LAB13 11482.2 7.12 1.50E−02 24 LAB13 11482.2 8.91 4.7E−02 42LAB41 11554.3 6.74 1.68E−02 17 LAB41 11551.2 8.47 3.3E−02 35 LAB2311572.6 7.38 2.39E−04 28 LAB41 11554.3 9.09 1.6E−02 45 Control 5.77LAB23 11572.6 12.36 5.5E−05 96 LAB11 11024.4 6.90 1.01E−02 22 Control6.29 0.00 LAB22 11064.6 6.46 1.27E−02 14 LAB11 11024.4 9.65 4.3E−02 54Control 5.66 Control 6.28 0.00 LAB32 11162.2 7.16 5.30E−03 16 LAB3811434.4 8.50 4.5E−02 30 LAB34 11171.4 6.80 4.04E−02 11 Control 6.55 0.00LAB38 11434.4 6.82 3.42E−02 11 LAB4 11962.1 6.17 2.4E−02 20 Control 6.14LAB4 11964.2 6.63 2.3E−02 29 LAB20 11131.1 6.50 2.83E−02 17 Control 5.130.00 LAB20 11132.7 6.63 3.94E−03 19 LAB20 11131.1 7.66 3.0E−02 38 LAB912284.1 6.31 1.42E−02 13 LAB9 12284.1 9.08 6.5E−03 64 LAB45 12361.1 6.601.08E−02 19 LAB9 12286.1 8.19 8.4E−03 48 LAB45 12363.2 6.81 5.63E−03 23LAB45 12361.1 9.06 1.3E−02 63 LAB45 12364.2 6.56 1.99E−02 18 LAB4512361.2 8.20 3.5E−02 48 LAB45 12365.1 7.02 5.96E−03 26 LAB45 12363.27.28 1.0E−02 31 LAB8 12423.3 6.28 3.68E−02 13 LAB45 12365.1 11.211.5E−03 102 LAB8 12425.4 6.88 5.67E−04 24 LAB8 12423.3 7.43 2.9E−03 34Control 5.56 LAB8 12425.4 10.37 1.3E−02 87 LAB32 11162.2 6.17 4.14E−0325 Control 5.55 0.00 LAB31 11421.5 5.74 2.02E−02 16 LAB20 11131.1 7.329.7E−03 42 LAB31 11423.4 5.66 4.38E−02 14 LAB20 11131.2 7.28 7.5E−03 42LAB13 11482.2 6.19 3.14E−03 25 LAB24 11191.5 6.51 4.7E−02 27 LAB812423.4 5.74 3.33E−02 16 LAB24 11193.5 6.62 4.5E−02 29 Control 4.95 0.00LAB49 11281.2 6.48 3.6E−02 26 LAB20 11131.1 6.09 1.16E−02 20 LAB4911283.5 8.10 1.4E−02 58 LAB20 11131.2 6.81 9.93E−03 34 LAB3 11331.1 6.711.7E−02 30 LAB20 11132.7 5.97 7.15E−04 18 Control 5.14 0.00 LAB2011134.4 5.81 1.05E−02 14 LAB24 11191.5 6.43 7.00E−05 27 LAB24 11192.16.15 3.99E−03 21 LAB24 11193.5 6.55 2.20E−04 29 LAB24 11193.6 6.003.51E−02 18 LAB49 11281.2 6.09 5.70E−05 20 LAB49 11281.4 6.29 9.66E−0324 LAB49 11283.1 6.01 2.98E−03 18 LAB49 11283.5 6.69 4.50E−03 32 LAB311331.1 6.50 4.00E−06 28 LAB3 11333.1 6.55 2.33E−04 29 LAB3 11333.5 6.051.00E−04 19 LAB3 11334.1 5.83 2.89E−02 15 LAB5 11443.3 6.07 1.28E−04 20LAB5 11444.1 5.99 7.11E−04 18 LAB36 11583.1 6.02 1.37E−02 19 LAB3611584.5 6.23 4.30E−05 23 LAB36 11585.5 6.74 3.65E−03 33 Control 5.08LAB2 11234.2 5.79 0.025 15.69% LAB2 11231.1 5.93 0.007 18.61% Control 5LAB2 11231.1 5.8 0.023 13.73% Control 5.1 Table 12: Analyses of plantroots (root length and root coverage) of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (At6669; SEQ ID NO: 674)when grown under limiting nitrogen conditions [low nitrogen or nitrogendeficient conditions (0.75 mMN)] as compared to control plants. “Incr.”= increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 13 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass undernitrogen deficient conditions Gene Plant Fresh Weight [g] Gene Plant DryWeight [g] Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. CTF163 11011.2 0.10 0.00 CTF215 11072.1 0.0089 1.5E−03 53 Control0.14 4.1E−02 31 Control 0.0058 CTF215 11072.1 0.16 2.1E−02 27 Control0.13 0.00 CTF226 10982.1 0.20 1.0E−03 49 CTF226 10982.3 0.18 4.7E−02 40Control 0.13 0.00 Table 13: Analyses of plant Biomass (fresh weight anddry weight) of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S; SEQ ID NO: 675) when grown under limitingnitrogen conditions [low nitrogen or nitrogen deficient conditions (0.75mMN)] as compared to control plants. “Incr.” = increment with respect toa control plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

TABLE 14 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass undernitrogen deficient conditions Gene Plant Fresh Weight [g] Gene Plant DryWeight [g] Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. LAB31 11421.5 0.130 1.9E−03 62 LAB31 11423.4 0.0057 3.0E−05 39LAB31 11423.4 0.125 2.6E−04 55 LAB41 11554.3 0.0062 6.8E−03 52 LAB1311482.2 0.119 8.8E−03 48 LAB23 11572.6 0.0079 8.3E−04 93 LAB41 11554.30.119 7.0E−03 48 LAB23 11573.4 0.0051 3.8E−03 24 LAB23 11572.6 0.1643.7E−03 105 LAB23 11574.2 0.0051 3.7E−02 26 Control 0.080 0.00 Control0.0041 0.00 LAB11 11022.3 0.142 4.0E−02 83 LAB11 11024.4 0.0069 2.3E−0481 LAB11 11024.4 0.155 4.6E−04 99 LAB16 11032.2 0.0048 2.9E−02 25Control 0.078 0.00 Control 0.0038 0.00 LAB15 11642.2 0.107 4.0E−02 39LAB32 11163.2 0.0061 2.0E−02 67 Control 0.077 0.00 LAB25 11341.2 0.00473.8E−02 28 LAB18 11653.4 0.087 3.5E−02 27 LAB38 11434.4 0.0045 2.8E−0224 Control 0.068 0.00 LAB15 11642.2 0.0053 2.4E−04 45 LAB20 11131.20.120 6.5E−04 85 Control 0.0037 0.00 LAB9 12281.2 0.100 1.1E−02 55 LAB1811653.4 0.0042 3.1E−02 19 LAB9 12286.1 0.108 4.8E−02 67 Control 0.00350.00 LAB45 12361.2 0.088 2.6E−03 35 LAB20 11131.2 0.0056 9.5E−04 91Control 0.065 0.00 Control 0.0029 0.00 LAB32 11162.2 0.107 8.4E−03 44LAB51 11561.5 0.0070 2.9E−02 92 LAB31 11423.4 0.119 3.0E−02 60 LAB812423.1 0.0049 5.0E−02 36 LAB13 11484.2 0.110 8.7E−03 47 Control 0.00360.00 LAB51 11561.5 0.139 3.6E−03 86 LAB49 11283.5 0.0054 8.3E−03 52LAB51 11563.2 0.105 3.8E−02 41 LAB5 11444.5 0.0059 2.5E−03 65 Control0.075 0.00 LAB36 11585.5 0.0052 8.7E−05 45 LAB20 11131.1 0.095 4.8E−0229 Control 0.0036 0.00 LAB20 11134.4 0.098 2.2E−02 33 LAB49 11283.50.109 6.3E−04 48 LAB3 11333.5 0.094 1.5E−02 28 LAB3 11334.1 0.0962.5E−02 30 LAB36 11583.1 0.101 4.0E−03 37 LAB36 11584.5 0.090 3.5E−02 23LAB36 11585.5 0.117 2.1E−03 59 Control 0.073 0.00 Table 14: Analyses ofplant Biomass (fresh weight and dry weight) of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (At6669; SEQ ID NO: 674)when grown under limiting nitrogen conditions [low nitrogen or nitrogendeficient conditions (0.75 mMN)] as compared to control plants. “Incr.”= increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 15 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass and growthrate under nitrogen deficient conditions Relative growth rate ofRelative growth rate of root root length (regression coverage(regression Gene coefficient) Gene coefficient) Name Event # Ave.p-value % incr. Name Event # Ave. p-value % incr. BDL103 8033.1 0.5712.2E−03 28.0% BDL103 8033.1 0.96 2.2E−03 38 BDL103 8033.4 0.543 8.5E−0321.9% Control 0.70 0.0 Control 0.446 0.0% CTF163 11011.2 0.74 2.0E−02 33CTF163 11012.2 0.525 5.0E−04 28.2% Control 0.55 0.0 CTF163 11012.7 0.5175.3E−03 26.2% CTF163 11011.2 0.84 1.8E−03 48 CTF215 11073.4 0.5071.9E−02 23.7% CTF163 11012.2 0.91 1.4E−03 61 Control 0.410 0.0% CTF16311012.7 0.95 2.9E−05 67 CTF163 11011.2 0.542 5.7E−04 43.0% Control 0.570.0 CTF163 11012.2 0.573 2.2E−04 51.2% CTF226 10982.1 1.06 6.7E−05 68CTF163 11012.4 0.503 1.7E−02 32.8% CTF226 10982.3 1.25 1.0E−06 99 CTF16311012.7 0.572 1.2E−04 50.9% CTF226 10985.1 0.80 4.9E−02 28 Control 0.3790.0% Control 0.63 0.0 CTF226 10982.3 0.646 1.5E−04 34.8% CTF205 11972.30.76 1.2E−03 37 Control 0.480 0.0% Control 0.56 0.0 Table 15: Analysesof root growth rate (relative growth rate of root length and rootcoverage) of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S; SEQ ID NO: 675) when grown under limitingnitrogen conditions [low nitrogen or nitrogen deficient conditions (0.75mM N)] as compared to control plants. “Incr.” = increment with respectto a control plant which has been transformed with an empty vector. Ave.= Average calculated from several transgenic events. “Event #” = numberof event (transgenic transformation).

TABLE 16 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass andgrowth rate under nitrogen deficient conditions Relative growth rate ofRelative growth rate of root root length (Regression coverage(regression Gene coefficient) Gene coefficient) Name Event # Ave.p-value % incr. Name Event # Ave. p-value % incr. LAB31 11422.1 0.582.8E−02 24 LAB31 11421.5 1.01 3.0E−02 42 LAB31 11423.4 0.62 1.3E−03 31LAB31 11423.4 1.09 2.8E−03 54 LAB13 11482.2 0.58 2.5E−02 24 LAB1311482.2 1.05 1.2E−02 49 LAB41 11551.2 0.59 4.0E−02 26 LAB41 11551.2 1.002.4E−02 41 LAB23 11572.6 0.62 4.0E−03 31 LAB41 11554.3 1.07 5.6E−03 51Control 0.47 0.0 LAB23 11572.6 1.45 1.0E−06 105 LAB32 11162.2 0.636.1E−03 22 Control 0.71 0.0 LAB34 11171.4 0.62 6.5E−03 22 LAB11 11024.41.13 6.0E−03 59 LAB38 11434.4 0.62 9.8E−03 20 Control 0.71 0.0 Control0.51 0.0 LAB32 11162.2 1.00 5.1E−03 36 LAB4 11964.2 0.57 2.6E−02 28LAB34 11171.4 0.96 7.1E−03 30 Control 0.44 0.0 LAB38 11434.4 1.013.4E−03 36 LAB20 11131.1 0.58 4.8E−02 27 LAB15 11642.2 1.05 5.2E−03 42LAB45 12365.1 0.60 4.0E−02 30 LAB15 11644.1 0.96 8.0E−03 30 LAB8 12425.40.59 3.7E−02 28 Control 0.74 0.0 Control 0.46 0.0 LAB4 11964.2 0.854.3E−02 41 LAB20 11131.2 0.56 1.4E−04 42 Control 0.60 0.0 LAB20 11132.10.46 3.5E−02 17 LAB4 11964.2 0.75 2.9E−02 32 LAB24 11191.5 0.51 1.7E−0431 Control 0.56 0.0 LAB24 11193.5 0.48 3.3E−03 22 LAB20 11131.1 0.883.5E−03 39 LAB24 11193.6 0.49 4.4E−03 24 LAB20 11132.7 0.81 3.7E−02 27LAB49 11281.2 0.50 7.1E−04 26 LAB9 12284.1 1.06 1.7E−05 68 LAB49 11283.50.57 1.7E−05 46 LAB9 12286.1 0.96 2.4E−04 51 LAB3 11331.1 0.52 1.0E−0533 LAB45 12361.1 1.05 4.6E−05 66 LAB3 11333.1 0.49 9.0E−04 25 LAB4512361.2 0.95 1.3E−03 50 LAB3 11334.1 0.45 4.4E−02 15 LAB45 12363.2 0.831.2E−02 30 LAB5 11443.3 0.45 5.0E−02 13 LAB45 12364.2 1.16 2.7E−04 83LAB5 11444.1 0.53 1.4E−05 34 LAB45 12365.1 1.30 0.0E+00 105 LAB5 11444.50.48 6.6E−03 22 LAB8 12423.3 0.88 2.0E−03 39 LAB36 11584.5 0.50 2.7E−0427 LAB8 12425.4 1.22 2.0E−06 92 LAB36 11585.5 0.53 1.2E−04 34 Control0.63 0.0 Control 0.39 0.0 LAB20 11131.1 0.80 1.5E−02 41 LAB20 11131.20.85 3.4E−03 49 LAB24 11191.5 0.74 4.9E−02 30 LAB24 11193.6 0.76 4.7E−0234 LAB49 11281.2 0.74 4.0E−02 31 LAB49 11283.5 0.97 4.0E−04 72 LAB311331.1 0.76 3.1E−02 34 LAB36 11584.5 0.74 4.3E−02 31 LAB36 11585.5 0.851.2E−02 51 Control 0.57 0.0 Table 16: Analyses of root growth rate(relative growth rate of root length and root coverage) of transgenicplants overexpressing the exogenous polynucleotides of some embodimentsof the invention (using the cloned or synthetic genes listed in Table 10above) under the regulation of a constitutive promoter (At6669; SEQ IDNO: 674) when grown under limiting nitrogen conditions [low nitrogen ornitrogen deficient conditions (0.75 mM N)] as compared to controlplants. “Incr.” = increment with respect to a control plant which hasbeen transformed with an empty vector. Ave. = Average calculated fromseveral transgenic events. “Event #” = number of event (transgenictransformation).

TABLE 17 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass and growthrate under nitrogen deficient conditions Relative growth rate of GeneLeaf area (cm²) Gene Leaf area Name Event # Ave. p-value % incr. NameEvent # Ave. p-value % incr. CTF113 5871.2 0.70 3.6E−02 21.6% CTF16311011.2 0.07 1.7E−02 24 Control 0.58 0.0% Control 0.06 0.00 CTF16311011.2 0.77 3.0E−02 28.4% CTF215 11072.1 0.08 4.6E−02 30 Control 0.600.0% Control 0.06 0.00 CTF215 11072.1 0.83 1.8E−02 27.9% CTF226 10982.10.11 1.1E−03 47 Control 0.65 0.0% CTF226 10982.3 0.11 7.9E−03 45 CTF22610982.1 1.16 8.3E−04 44.1% Control 0.08 0.00 CTF226 10982.3 1.10 1.6E−0236.4% Control 0.80 0.0% CTF180 11371.1 0.59 4.2E−02 16.2% Control 0.510.0% Table 17: Analyses of leaf area and leaf area growth rate (relativegrowth rate of leaf area) of transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S; SEQ ID NO: 675) when grownunder limiting nitrogen conditions [low nitrogen or nitrogen deficientconditions (0.75 mM N)] as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 18 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass andgrowth rate under nitrogen deficient conditions Relative growth rate ofleaf area (regression Gene Leaf area (cm²) Gene coefficient) Name Event# Ave. p-value % incr. Name Event # Ave. p-value % incr. LAB31 11421.50.79 4.9E−04 51.0% LAB31 11421.5 0.08 1.0E−05 53.6% LAB31 11423.4 0.692.4E−03 32.1% LAB31 11423.4 0.07 2.7E−05 45.4% LAB23 11572.6 0.937.0E−04 76.7% LAB13 11482.2 0.06 2.1E−02 23.5% Control 0.52 0.0% LAB1311484.2 0.06 1.7E−02 25.0% LAB11 11024.4 0.80 3.3E−05 48.2% LAB4111551.2 0.07 2.0E−03 34.5% Control 0.54 0.0% LAB41 11554.3 0.07 9.9E−0449.2% LAB32 11163.2 0.67 5.2E−03 17.9% LAB23 11572.6 0.09 0.0E+00 86.1%LAB17 11534.1 0.84 8.6E−03 48.4% Control 0.05 0.0% LAB15 11642.2 0.685.3E−03 20.5% LAB11 11024.4 0.08 4.4E−03 39.2% Control 0.57 0.0% Control0.06 0.0% LAB4 11964.2 0.52 4.0E−02 30.3% LAB25 11341.1 0.07 4.9E−0224.6% Control 0.40 0.0% LAB17 11534.1 0.08 1.8E−03 41.4% LAB20 11131.20.51 3.1E−04 43.7% Control 0.06 0.0% LAB9 12281.2 0.45 2.2E−02 28.1%LAB20 11131.2 0.05 1.3E−03 37.3% Control 0.35 0.0% LAB9 12281.2 0.051.9E−02 27.4% LAB20 11131.1 0.64 4.3E−03 46.9% LAB9 12284.1 0.05 2.6E−0248.5% LAB20 11134.4 0.53 2.5E−02 23.6% LAB9 12286.1 0.05 3.9E−02 30.3%LAB9 12284.1 0.79 1.8E−03 83.2% LAB45 12364.2 0.05 4.7E−02 30.6% LAB912286.1 0.56 4.3E−02 29.9% Control 0.04 0.0% LAB45 12365.1 0.75 2.5E−0272.8% LAB20 11131.1 0.07 5.9E−04 47.5% LAB8 12425.4 0.68 5.9E−03 56.7%LAB9 12284.1 0.08 1.0E−06 83.1% Control 0.43 0.0% LAB9 12286.1 0.061.3E−02 32.8% LAB31 11423.4 0.60 6.8E−04 49.2% LAB45 12361.1 0.064.5E−03 44.4% LAB13 11481.5 0.57 2.1E−02 43.5% LAB45 12365.1 0.083.8E−04 69.1% LAB51 11561.2 0.55 2.5E−02 36.9% LAB8 12422.3 0.06 2.6E−0237.8% LAB51 11561.5 0.55 3.3E−02 37.4% LAB8 12425.4 0.07 8.1E−05 58.7%Control 0.40 0.0% Control 0.04 0.0% LAB20 11131.1 0.55 2.1E−02 28.2%LAB31 11423.4 0.06 2.7E−02 35.4% LAB49 11283.5 0.58 3.7E−02 33.6%Control 0.04 0.0% LAB5 11444.5 0.54 2.6E−02 26.0% LAB49 11283.5 0.062.8E−02 41.4% Control 0.43 0.0% Control 0.04 0.0% Table 18: Analyses ofleaf area and leaf area growth rate (leaf area growth rate) oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter(At6669; SEQ ID NO: 674) when grown under limiting nitrogen conditions[low nitrogen or nitrogen deficient conditions (0.75 mM N)] as comparedto control plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

The genes presented in Tables 19-26, hereinbelow, were found to improveplant performance by improving root performance, plant growthcharacteristic and plant biomass when grown under normal growthconditions, compared to control plants.

Tables 19-26 depict analyses of root growth (root length and rootcoverage; Tables 19 and 20); plant biomass (plant fresh, dry weight andleaf area; Tables 21 and 22); root growth rate (relative growth rate ofroot length and root coverage; Tables 23 and 24); leaf area and leafarea growth rate (Relative growth rate of leaf area; Tables 25 and 26)when grown under normal growth conditions (i.e., in the presence of 15mM nitrogen) in plants overexpressing the polynucleotides of someembodiments of the invention under the regulation of a constitutivepromoter [35S (SEQ ID NO:675) or At6669 (SEQ ID NO:674)]. Evaluation ofeach gene was performed by testing the performance of several events.Some of the genes were evaluated in more than one tissue culture assayand the second experiment confirmed the significant increment in plantbiomass. Event with p-value <0.05 was considered statisticallysignificant.

TABLE 19 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant roots under normalconditions Gene Plant root length [cm] Gene Plant root coverage [cm²]Name Event # Ave. p-value % incr. Name Event # Ave. p-value % incr.BDL103 8033.4 5.29 7.6E−03 14.5% BDL166 9985.2 7.21 3.1E−02 67.0%Control 4.62 0.0% Control 4.32 0.0% CTF113 5871.3 2.51 2.7E−02 42.3%CTF163 11012.7 6.41 1.6E−02 40.2% Control 1.76 0.0% Control 4.58 0.0%CTF163 11012.7 6.38 9.6E−03 48.1% CTF205 11972.3 4.64 2.8E−02 47.5%Control 4.31 0.0% Control 3.15 0.0% CTF205 11972.3 5.26 5.5E−03 23.4%Control 4.26 0.0% Table 19: Analyses of plant roots (root length androot coverage) of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S; SEQ ID NO: 675) when grown under normalconditions as compared to control plants. “Incr.” = increment withrespect to a control plant which has been transformed with an emptyvector. Ave. = Average calculated from several transgenic events. “Event#” = number of event (transgenic transformation).

TABLE 20 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant roots undernormal conditions Gene Plant root length [cm] Gene Plant root coverage[cm²] Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. LAB31 11421.5 6.55 1.15E−02 21.9% LAB13 11482.2 8.17 1.1E−02 62.5%LAB31 11423.2 6.44 4.87E−02 20.0% LAB41 11551.2 6.90 2.7E−02 37.3% LAB3111423.4 6.32 4.20E−02 17.7% LAB23 11572.6 7.11 9.5E−03 41.5% LAB1311482.2 6.90 2.98E−03 28.4% Control 5.03 0.0% LAB13 11484.2 6.693.18E−02 24.5% LAB22 11062.3 5.77 7.7E−04 55.9% LAB41 11551.2 6.892.57E−03 28.4% LAB36 11585.5 5.21 6.3E−03 40.8% LAB23 11572.6 6.724.99E−03 25.2% Control 3.70 0.0% Control 5.37 0.0% LAB34 11175.1 5.976.2E−03 50.8% LAB11 11022.1 5.79 8.49E−03 22.2% LAB50 Control 4.86 0.0%LAB11 11022.3 6.25 1.42E−03 31.9% LAB4 11964.2 4.70 4.1E−02 41.8% LAB1111024.4 5.87 1.96E−02 23.9% LAB18 Control 3.31 0.0% LAB16 11032.2 5.841.06E−02 23.4% LAB4 11964.1 5.30 1.5E−02 30.7% LAB22 11062.3 6.092.15E−03 28.6% Control 4.05 0.0% LAB22 11063.4 5.66 2.93E−02 19.6% LAB912281.2 5.45 2.5E−02 55.1% LAB22 11064.6 5.78 1.99E−02 21.9% LAB4512365.1 5.91 3.5E−02 68.1% LAB36 11585.5 5.77 3.64E−03 21.8% Control3.51 0.0% Control 4.74 0.0% LAB9 12284.1 8.02 6.4E−03 69.3% LAB3211162.2 5.27 2.10E−02 14.2% LAB45 12365.1 7.92 1.8E−03 67.2% LAB3211163.2 5.91 2.14E−02 27.9% LAB8 12423.1 7.10 6.2E−03 50.0% LAB3411175.1 5.55 1.21E−02 20.1% LAB8 12425.4 7.47 3.8E−03 57.8% LAB3311272.4 5.07 4.85E−02 9.8% Control 4.74 0.0% LAB25 11341.1 5.79 3.40E−0225.4% LAB49 11281.4 5.43 3.3E−02 26.9% LAB15 11642.2 5.74 3.61E−02 24.2%Control 4.28 0.0% LAB15 11644.2 5.15 3.29E−02 11.6% Control 4.62 0.0%LAB2 11234.2 5.18 4.51E−02 20.9% LAB4 11962.1 5.15 2.22E−02 20.4% LAB411963.2 4.98 1.41E−02 16.3% Control 4.28 0.0% LAB20 11131.1 5.523.31E−02 20.6% LAB9 12281.2 5.22 6.31E−03 14.1% LAB45 12365.1 5.841.27E−03 27.6% Control 4.58 0.0% LAB45 12361.1 6.10 2.44E−02 11.2% LAB812425.4 6.31 8.97E−03 15.0% Control 5.48 0.0% LAB13 11482.2 5.864.74E−03 21.5% Control 4.82 0.0% LAB20 11131.2 6.81 8.98E−04 28.8% LAB2011132.7 6.58 1.90E−02 24.6% LAB20 11134.4 6.12 1.60E−02 15.8% LAB2411193.5 6.10 2.51E−03 15.6% LAB3 11333.1 6.45 1.02E−02 22.2% LAB3611584.5 6.53 3.45E−02 23.6% Control 5.28 0.0% LAB2 11234.2 5.17 0.0457.81% Control 4.80 Table 20: Analyses of plant roots (root length androot coverage) of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (At6669; SEQ ID NO: 674) when grown under normalconditions as compared to control plants. “Incr.” = increment withrespect to a control plant which has been transformed with an emptyvector. Ave. = Average calculated from several transgenic events. “Event#” = number of event (transgenic transformation).

TABLE 21 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass undernormal conditions Gene Plant Fresh Weight [g] Gene Plant Dry Weight [g]Name Event # Ave. p-value % incr. Name Event # Ave. p-value % incr.CTF113 5872.1 0.15 2.1E−02 44.4% CTF113 5872.1 0.0086 2.3E−02 0.44Control 0.10 0.0% Control Control 0.0060 0.00 BDL14 5761.2 0.24 3.3E−0244.5% CTF175 8701.4 0.0054 4.5E−03 0.91 Control 0.16 0.0% CTF175 8702.40.0042 4.9E−02 0.49 BDL17 6081.3 0.30 2.9E−02 63.5% Control Control0.0028 0.00 Control 0.18 0.0% BDL103 8033.12 0.0050 8.3E−03 0.13 BDL1669985.2 0.29 5.9E−03 52.2% Control Control 0.0107 0.00 Control 0.19 0.0%BDL166 9985.2 0.0163 8.5E−04 0.73 CTF226 10985.1 0.19 3.1E−02 40.0%Control Control 0.0094 0.00 CTF226 10985.5 0.17 2.2E−02 29.7% CTF21511072.1 0.0080 2.6E−02 0.29 Control 0.13 0.0% Control Control 0.00620.00 CTF205 11972.3 0.11 1.2E−02 32.1% CTF226 10982.1 0.0097 2.5E−030.69 Control 0.09 0.0% CTF226 10985.1 0.0081 1.5E−02 0.42 BDL17 6081.30.20 4.7E−03 78.5% Control Control 0.0057 0.00 BDL17 6081.5 0.20 2.2E−0283.1% CTF205 11972.3 0.0053 5.0E−03 0.20 BDL17 6083.2 0.21 1.0E−04 94.9%Control Control 0.0044 0.00 Control 0.11 0.0% BDL17 6081.3 0.00884.0E−02 0.49 CTF180 11371.1 0.11 5.7E−04 63.2% BDL17 6083.2 0.01062.1E−02 0.79 CTF180 11376.1 0.09 1.3E−02 29.5% Control Control 0.00590.00 CTF205 11972.3 0.10 1.6E−02 46.2% CTF180 11371.1 0.0044 5.0E−020.49 CTF205 11973.2 0.09 3.7E−02 35.0% CTF180 11376.1 0.0038 4.0E−020.28 Control 0.07 0.0% Control Control 0.0030 0.00 Table 21: Analyses ofplant Biomass (fresh weight and dry weight) of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S; SEQ ID NO: 675)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 22 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass undernormal conditions Plant Fresh Weight [g] Plant Dry Weight [g] Gene GeneName Event # Ave. p-value % incr. Name Event # Ave. p-value % incr.LAB23 11572.6 0.129 4.2E−02 31.2% LAB23 11571.5 0.0057 1.0E−02 0.30Control 0.098 0.0% LAB23 11572.6 0.0062 1.1E−03 0.43 LAB11 11022.1 0.1023.2E−03 43.8% Control Control 0.0044 0.00 LAB11 11022.3 0.099 5.7E−0339.2% LAB11 11022.1 0.0045 3.4E−02 0.53 LAB16 11032.2 0.126 1.5E−0277.4% LAB11 11023.4 0.0035 4.9E−02 0.18 LAB16 11033.2 0.098 2.1E−0237.9% LAB11 11024.4 0.0044 3.6E−02 0.52 LAB22 11064.6 0.125 3.7E−0275.7% LAB22 11064.6 0.0055 1.4E−02 0.89 LAB36 11585.5 0.091 1.7E−0227.3% LAB36 11584.2 0.0042 3.7E−02 0.43 Control 0.071 0.0% LAB36 11584.50.0036 8.8E−03 0.24 LAB32 11163.2 0.116 8.2E−03 39.8% Control Control0.0029 0.00 LAB33 11272.2 0.120 6.0E−03 43.7% LAB32 11163.2 0.00571.9E−02 0.40 LAB17 11534.1 0.110 4.6E−02 31.9% LAB33 11272.2 0.00511.9E−02 0.25 LAB15 11642.2 0.154 2.8E−02 85.2% LAB15 Control 0.0041 0.00Control 0.083 0.0% LAB18 11653.7 0.0037 2.7E−02 0.24 LAB18 11653.7 0.0763.3E−02 26.9% Control Control 0.0030 0.00 Control 0.060 0.0% LAB2011131.2 0.0063 3.8E−02 1.24 LAB20 11131.2 0.167 1.4E−04 146.4% LAB912282.2 0.0039 2.4E−02 0.37 LAB20 11134.4 0.095 2.1E−02 40.7% LAB912284.1 0.0057 4.2E−02 1.01 LAB9 12284.1 0.133 4.7E−02 95.8% LAB4512365.1 0.0047 2.6E−02 0.66 Control 0.068 0.0% Control Control 0.00280.00 LAB32 11163.1 0.089 3.8E−03 40.3% LAB9 12281.2 0.0063 2.2E−02 0.30LAB31 11422.5 0.099 1.4E−02 55.8% LAB9 12284.1 0.0077 1.5E−03 0.58 LAB3111423.1 0.106 8.6E−03 66.6% Control Control 0.0049 0.00 LAB13 11481.50.102 8.7E−05 60.6% LAB32 11163.1 0.0046 9.9E−04 0.67 LAB13 11482.20.106 2.2E−02 65.9% LAB31 11422.1 0.0041 2.5E−02 0.48 LAB13 11483.20.114 4.3E−02 79.4% LAB31 11423.1 0.0050 4.3E−02 0.82 LAB13 11483.30.081 4.5E−02 27.2% LAB31 11423.4 0.0053 3.5E−02 0.91 LAB13 11484.20.127 5.0E−04 99.6% LAB13 11483.2 0.0051 3.2E−02 0.85 LAB51 11561.20.107 8.1E−03 67.3% LAB13 11484.2 0.0063 8.4E−03 1.27 LAB51 11561.50.135 2.7E−02 112.1% LAB51 11561.2 0.0039 3.5E−02 0.42 LAB51 11564.70.119 3.3E−02 86.9% Control Control 0.0028 0.00 LAB8 12422.3 0.0793.6E−02 24.1% LAB24 11193.6 0.0041 6.2E−03 0.44 LAB8 12423.3 0.0812.5E−02 27.5% LAB3 11334.1 0.0042 3.4E−03 0.47 Control 0.064 0.0% LAB511443.4 0.0039 4.8E−03 0.36 LAB3 11333.5 0.128 1.6E−03 53.0% LAB511444.5 0.0044 1.7E−02 0.54 Control 0.084 0.0% LAB36 11583.1 0.00449.1E−04 0.57 Control    .5 0.0028 0.00 Table 22: Analyses of plantbiomass (fresh weight and dry weight) of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (At6669; SEQ ID NO: 674)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 23 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass and growthrate under normal conditions Relative growth rate of root lengthRelative growth rate of root coverage (regression coefficient)(regression coefficient) Gene Gene Name Event # Ave. p-value % incr.Name Event # Ave. p-value % incr. BDL103 8033.4 0.509 2.23E−02 21.9%CTF113 5871.3 0.27 2.3E−03 165.9% Control 0.418 0.0% Control 0.10 0.0%CTF113 5871.3 0.192 3.31E−02 49.6% BDL210 10831.5 0.42 4.9E−02 54.4%Control 0.128 0.0% Control 0.27 0.0% BDL210 10831.5 0.356 3.29E−02 36.5%BDL166 9985.2 0.83 1.2E−02 62.4% Control 0.261 0.0% Control 0.51 0.0%CTF163 11012.7 0.604 8.70E−05 64.9% CTF163 11012.7 0.75 4.0E−03 42.6%Control 0.366 0.0% Control 0.53 0.0% BDL17 6081.5 1.12 3.2E−02 52.6%Control 0.74 0.0% CTF205 11972.3 0.50 1.0E−02 38.7% Control 0.36 0.0%Table 23: Analyses of root growth rate (relative growth rate of rootlength and root coverage) of transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S; SEQ ID NO: 675) when grownunder normal conditions as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 24 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass andgrowth rate under normal conditions Relative Growth Rate of Root LengthRelative Growth Rate of Root Coverage (Regression coefficient)(Regression coefficient) Gene Gene Name Event # Ave. p-value % incr.Name Event # Ave. p-value % incr. LAB13 11482.2 0.594 1.86E−02 38.3%LAB31 11423.4 0.83 1.6E−02 52.0% LAB41 11551.2 0.618 6.18E−03 43.8%LAB13 11482.2 0.97 4.6E−04 79.0% Control 0.430 0.0% LAB13 11484.2 0.792.6E−02 45.1% LAB22 11062.3 0.498 1.22E−02 31.6% LAB41 11551.2 0.801.7E−02 47.3% LAB36 11585.5 0.487 1.84E−02 28.8% LAB41 11552.4 0.774.7E−02 42.4% Control 0.378 0.0% LAB41 11554.3 0.94 4.9E−03 73.6% LAB2111144.1 0.423 2.99E−02 25.1% LAB23 11572.6 0.82 7.6E−03 51.2% LAB3211162.2 0.412 3.64E−02 21.9% Control 0.54 0.0% LAB32 11163.2 0.4551.59E−02 34.7% LAB11 11022.1 0.59 1.7E−02 45.8% LAB34 11175.1 0.4321.57E−02 27.8% LAB11 11022.3 0.58 1.4E−02 42.6% LAB25 11341.1 0.4578.49E−03 35.1% LAB16 11032.3 0.56 2.1E−02 38.3% LAB25 11342.2 0.4643.22E−03 37.3% LAB22 11062.3 0.67 2.1E−04 65.5% LAB38 11434.4 0.4312.27E−02 27.6% LAB36 11585.5 0.59 6.3E−03 45.2% LAB15 11641.1 0.4802.38E−03 42.0% Control 0.41 0.0% LAB15 11642.2 0.464 7.31E−03 37.3%LAB32 11163.2 0.60 6.9E−03 46.9% Control 0.338 0.0% LAB34 11175.1 0.648.6E−04 56.7% LAB2 11231.1 0.430 4.51E−02 24.2% LAB25 11341.1 0.553.7E−02 34.9% LAB4 11962.1 0.439 1.79E−02 26.6% LAB25 11342.2 0.593.3E−02 44.3% LAB4 11964.2 0.480 5.38E−03 38.6% LAB17 11533.7 0.574.9E−02 39.4% Control 0.346 0.0% LAB15 11642.2 0.73 1.5E−03 78.2% LAB2011131.1 0.476 3.64E−03 32.5% LAB15 11644.1 0.58 2.1E−02 41.2% LAB4512364.2 0.439 4.72E−02 22.2% LAB15 11644.2 0.58 1.4E−02 42.7% LAB4512365.1 0.462 5.73E−03 28.6% Control 0.41 0.0% Control 0.359 0.0% LAB411964.2 0.55 3.0E−03 48.5% LAB13 11482.2 0.505 2.79E−02 26.0% Control0.37 0.0% Control 0.401 0.0% LAB2 11231.1 0.58 2.6E−02 39.5% LAB2011131.2 0.543 1.00E−05 40.0% LAB4 11963.2 0.65 4.0E−02 54.6% LAB2011132.7 0.527 1.08E−03 35.9% LAB4 11964.1 0.58 1.8E−02 37.8% LAB4911281.4 0.482 3.02E−02 24.3% Control 0.42 0.0% LAB3 11333.1 0.5194.80E−04 33.7% LAB20 11131.1 0.49 3.3E−02 26.5% LAB5 11444.1 0.4709.27E−03 21.3% LAB20 11131.2 0.55 2.9E−02 41.2% LAB36 11584.2 0.4851.39E−02 25.0% LAB9 12281.2 0.60 4.2E−04 55.9% LAB36 11584.5 0.5462.23E−04 40.7% LAB45 12361.2 0.62 7.1E−03 61.2% Control 0.388 0.0% LAB4512364.2 0.54 8.5E−03 39.6% LAB2 11231.1 0.43 0.04 LAB45 12365.1 0.638.7E−04 63.0% Control 0.35 20.16% Control 0.39 0.0% LAB9 12284.1 0.899.8E−04 68.3% LAB45 12361.2 0.75 2.9E−02 42.1% LAB45 12365.1 0.912.0E−04 71.0% LAB8 12423.1 0.80 5.6E−03 50.6% LAB8 12425.4 0.81 3.3E−0353.1% Control 0.53 0.0% LAB13 11482.2 0.56 2.5E−02 47.4% LAB51 11561.50.63 1.3E−02 64.1% Control 0.38 0.0% LAB20 11131.2 0.58 4.1E−02 27.6%LAB20 11132.7 0.61 2.1E−02 32.9% LAB49 11281.4 0.59 1.9E−02 28.8%Control 0.46 0.0% LAB2 11231.1 0.58 0.025 39.5% Control 0.41 Table 24:Analyses of root growth rate (relative growth rate of root length androot coverage) of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (At6669; SEQ ID NO: 674) when grown under normalconditions as compared to control plants. “Incr.” = increment withrespect to a control plant which has been transformed with an emptyvector. Ave. = Average calculated from several transgenic events. “Event#” = number of event (transgenic transformation).

TABLE 25 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass and growthrate under normal conditions Relative growth rate of leaf area Leaf area(cm²) (regression coefficient) Gene Gene Name Event # Average p-value %incr. Name Event # Average p-value % incr. BDL17 6081.3 0.80 3.7E−0335.7% CTF113 5873.3 0.06 1.5E−02 46.6% Control 0.59 0.0% Control 0.040.0% BDL103 8033.1 0.82 8.7E−03 39.5% BDL17 6081.3 0.08 3.7E−02 34.6%BDL103 8033.12 0.66 9.7E−04 12.3% Control 0.06 0.0% Control 1.16 0.0%BDL103 8033.1 0.09 3.2E−02 38.2% BDL166 9985.2 1.13 1.6E−03 40.7% BDL1038033.12 0.07 2.5E−03 8.1% Control 0.80 0.0% Control 0.13 0.0% CTF22610982.1 1.24 2.3E−02 37.6% BDL166 9985.2 0.11 4.8E−03 41.5% Control 0.900.0% Control 0.08 0.0% BDL17 6081.3 1.21 3.8E−03 54.7% CTF215 11072.10.09 3.3E−02 34.4% BDL17 6081.5 1.17 1.0E−02 50.0% Control 0.07 0.0%BDL17 6083.2 1.20 7.1E−03 54.0% CTF205 11972.3 0.06 3.7E−02 25.3%Control 0.78 0.0% Control 0.05 0.0% CTF180 11371.1 0.52 2.8E−02 39.5%BDL17 6081.3 0.11 2.2E−02 43.1% CTF180 11376.1 0.48 8.0E−03 27.8% BDL176081.5 0.12 1.5E−02 46.9% CTF205 11973.2 0.53 2.1E−02 41.3% BDL17 6083.20.12 2.8E−03 59.2% Control 0.38 0.0% Control 0.08 0.0% CTF180 11371.10.05 1.1E−02 39.3% CTF180 11376.1 0.05 4.0E−02 28.6% CTF205 11973.2 0.054.5E−03 44.8% Control 0.04 0.0% Table 25: Analyses of leaf area and leafarea growth rate (leaf area growth rate) of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S; SEQ ID NO: 675)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 26 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the At6669 promoter exhibit improved plant biomass andgrowth rate under normal conditions Leaf area (cm²) Relative growth rateof leaf area Gene Gene Name Event # Average p-value % incr. Name Event #Average p-value % incr. LAB41 11554.3 0.66 4.8E−02 29.3% LAB31 11421.50.08 1.0E−05 53.6% LAB23 11572.6 0.76 1.9E−02 47.9% LAB31 11423.4 0.072.7E−05 45.4% Control 0.51 0.0% LAB13 11482.2 0.06 2.1E−02 23.5% LAB1111022.3 0.60 5.0E−03 31.9% LAB13 11484.2 0.06 1.7E−02 25.0% LAB1111024.4 0.69 2.8E−04 50.7% LAB41 11551.2 0.07 2.0E−03 34.5% LAB1611032.2 0.62 4.4E−02 36.7% LAB41 11554.3 0.07 9.9E−04 49.2% LAB2211064.6 0.63 1.9E−02 38.5% LAB23 11572.6 0.09 0.0E+00 86.1% LAB3611584.2 0.61 2.2E−02 33.9% Control 0.05 0.0% Control 0.46 0.0% LAB1111024.4 0.08 4.4E−03 39.2% LAB33 11272.2 0.65 1.2E−02 32.4% Control 0.060.0% LAB25 11341.2 0.64 3.8E−02 30.4% LAB25 11341.1 0.07 4.9E−02 24.6%LAB17 11534.1 0.70 8.2E−05 42.1% LAB17 11534.1 0.08 1.8E−03 41.4% LAB1511642.2 0.82 4.6E−02 67.1% Control 0.06 0.0% Control 0.49 0.0% LAB2011131.2 0.05 1.3E−03 37.3% LAB20 11131.2 0.58 2.0E−02 88.7% LAB9 12281.20.05 1.9E−02 27.4% LAB9 12284.1 0.51 3.3E−02 65.6% LAB9 12284.1 0.052.6E−02 48.5% LAB45 12361.1 0.44 1.8E−03 45.1% LAB9 12286.1 0.05 3.9E−0230.3% Control 0.31 0.0% LAB45 12364.2 0.05 4.7E−02 30.6% LAB9 12284.10.85 1.3E−02 78.6% Control 0.04 0.0% LAB45 12365.1 0.83 1.3E−04 74.9%LAB20 11131.1 0.07 5.9E−04 47.5% LAB8 12423.1 0.62 2.9E−02 31.4% LAB912284.1 0.08 1.0E−06 83.1% Control 0.48 0.0% LAB9 12286.1 0.06 1.3E−0232.8% LAB32 11163.1 0.41 4.0E−02 26.3% LAB45 12361.1 0.06 4.5E−03 44.4%LAB31 11422.5 0.40 3.4E−02 25.1% LAB45 12365.1 0.08 3.8E−04 69.1% LAB3111423.1 0.48 4.0E−02 49.2% LAB8 12422.3 0.06 2.6E−02 37.8% LAB31 11423.40.55 3.3E−02 69.4% LAB8 12425.4 0.07 8.1E−05 58.7% LAB13 11481.5 0.544.3E−03 68.2% Control 0.04 0.0% LAB13 11482.2 0.43 1.5E−02 34.7% LAB3111423.4 0.06 2.7E−02 35.4% LAB13 11483.2 0.58 3.3E−03 80.1% Control 0.040.0% LAB13 11484.2 0.69 1.8E−02 114.1% LAB49 11283.5 0.06 2.8E−02 41.4%LAB51 11561.2 0.45 1.5E−02 41.2% Control 0.04 0.0% LAB51 11561.5 0.583.4E−02 81.4% LAB51 11563.2 0.42 1.2E−02 31.4% LAB51 11564.5 0.483.1E−02 47.9% LAB51 11564.7 0.53 1.0E−02 63.2% LAB8 12422.3 0.42 2.9E−0231.9% LAB8 12423.1 0.40 4.7E−02 25.5% Control 0.32 0.0% LAB20 11132.10.47 2.2E−02 19.9% LAB49 11281.4 0.50 2.4E−03 30.1% LAB3 11333.5 0.501.0E−03 28.8% LAB5 11444.5 0.58 1.3E−02 49.4% LAB36 11583.1 0.47 1.8E−0221.6% LAB36 11585.5 0.48 7.4E−03 22.9% Control 0.39 0.0% Table 26:Analyses of leaf area and leaf area growth rate (leaf area growth rate)of transgenic plants overexpressing the exogenous polynucleotides ofsome embodiments of the invention (using the cloned or synthetic geneslisted in Table 10 above) under the regulation of a constitutivepromoter (At6669; SEQ ID NO: 674) when grown under normal conditions ascompared to control plants. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

Example 7 Evaluation of Transgenic Arabidopsis Plant Growth UnderAbiotic Stress as Well as Under Favorable Conditions in

Greenhouse Assay Grown Until Seed Production

ABS Tolerance: Yield and Plant Growth Rate at High SalinityConcentration Under Greenhouse Conditions—

This assay follows the rosette area growth of plants grown in thegreenhouse as well as seed yield at high salinity irrigation. Seeds weresown in agar media supplemented only with a selection agent (Kanamycin)and Hoagland solution under nursery conditions. The T₂ transgenicseedlings were then transplanted to 1.7 trays filled with peat andperlite. The trays were irrigated with tap water (provided from thepots' bottom). Half of the plants were irrigated with a salt solution(40-80 mM NaCl and 5 mM CaCl₂) so as to induce salinity stress (stressconditions). The other half of the plants was irrigated with tap water(normal conditions). All plants were grown in the greenhouse untilmature seeds, then harvested (the above ground tissue) and weighted(immediately or following drying in oven at 50° C. for 24 hours). Highsalinity conditions were achieved by irrigating with a solutioncontaining 40-80 mM NaCl (“ABS” growth conditions) and compared toregular growth conditions.

Each construct was validated at its T₂ generation. Transgenic plantstransformed with a construct including the uidA reporter gene (GUS)under the At6669 promoter (SEQ ID NO:674) or with an empty vectorincluding the At6669 promoter are used as control.

The plants were analyzed for their overall size, growth rate, flowering,seed yield, weight of 1,000 seeds, dry matter and harvest index (HI-seedyield/dry matter). Transgenic plants performance was compared to controlplants grown in parallel under the same conditions. Mock-transgenicplants expressing the uidA reporter gene (GUS-Intron) or with no gene atall (empty vector, containing the Kan selection gene), under the samepromoter were used as control.

The experiments were planned in nested randomized plot distribution. Foreach gene of the invention three to five independent transformationevents were analyzed from each construct.

Digital Imaging—

A laboratory image acquisition system, which consists of a digitalreflex camera (Canon EOS 300D) attached with a 55 mm focal length lens(Canon EF-S series), mounted on a reproduction device (Kaiser RS), whichincluded 4 light units (4×150 Watts light bulb) was used for capturingimages of plant samples.

The image capturing process was repeated every 2 days starting from day1 after transplanting till day 16. Same camera, placed in a custom madeiron mount, was used for capturing images of larger plants sawn in whitetubs in an environmental controlled greenhouse. The tubs were squareshape and include 1.7 liter trays. During the capture process, the tubswere placed beneath the iron mount, while avoiding direct sun light andcasting of shadows.

An image analysis system was used, which consists of a personal desktopcomputer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ1.39 (Java based image processing program which was developed at theU.S. National Institutes of Health and freely available on the internetat Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Imageswere captured in resolution of 10 Mega Pixels (3888×2592 pixels) andstored in a low compression JPEG (Joint Photographic Experts Groupstandard) format. Next, analyzed data was saved to text files andprocessed using the JMP statistical analysis software (SAS institute).

Leaf Growth Analysis—

Using the digital analysis leaves data was calculated, including leafnumber, rosette area, rosette diameter, leaf blade area, plot coverage,leaf petiole length.

Vegetative growth rate: is the rate of growth of the plant as defined byformulas VIII, IX, XI and XI

Relative growth rate of leaf blade area=Regression coefficient of leafarea along time course.  Formula VIII:

Relative growth rate of rosette area=Regression coefficient of rosettearea along time course.  Formula IX:

Relative growth rate of rosette diameter=Regression coefficient ofrosette diameter along time course.  Formula X

Relative growth rate of plot coverage=Regression coefficient of plotcoverage along time course.  Formula XI

Seeds Average Weight (Seed Weight or 1000 Seed Weight)—

At the end of the experiment all seeds were collected. The seeds werescattered on a glass tray and a picture was taken. Using the digitalanalysis, the number of seeds in each sample was calculated.

Plant Dry Weight and Seed Yield—

On about day 80 from sowing, the plants were harvested and left to dryat 30° C. in a drying chamber. The biomass and seed weight of each plotwere measured and divided by the number of plants in each plot.

Dry weight=total weight of the vegetative portion above ground(excluding roots) after drying at 30° C. in a drying chamber;

Seed yield per plant=total seed weight per plant (grams).

The Harvest Index can be calculated using Formula IV (as describedabove; Harvest Index=Average seed yield per plant/Average dry weight).

Statistical Analyses—

To identify genes conferring significantly improved ABST, nitrogen useefficiency, growth rate, biomass, oil content and yield production, theresults obtained from the transgenic plants were compared to thoseobtained from control plants. To identify outperforming genes andconstructs, results from the independent transformation events testedwere analyzed separately. Data was analyzed using Student's t-test andresults were considered significant if the p value was less than 0.1.The JMP statistics software package was used (Version 5.2.1, SASInstitute Inc., Cary, N.C., USA).

Experiment Results:

The genes presented in Tables 27-31, hereinbelow, have improved plantABST when grown at high salinity irrigation levels (80-100 mM NaCl).These genes produced higher seed yield, harvest index, seed weight(expressed as 1000-seed weight) and plant biomass [(as expressed asplant dry weight (DW)] when grown under high salinity irrigationconditions, compared to control.

Tables 27-29 depict analyses of seed yield and weight (Table 27),harvest index (Table 28) and dry weight (Table 29) when grown under highsalinity irrigation conditions in plants overexpressing thepolynucleotides of some embodiments of the invention under theregulation of a constitutive (35S; SEQ ID NO:675). Evaluation of eachgene was performed by testing the performance of several events. Some ofthe genes were evaluated in more than one tissue culture assay and theresults obtained were repeated. Event with p-value <0.05 was consideredstatistically significant.

TABLE 27 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved seed yield and weightunder high salinity irrigation growth conditions Gene Seed yield GeneSeeds weight (g) Name Event # Ave. p-value % incr. Name Event # Ave.p-value % incr. CTF113 5871.2 0.03 3.0E−03 75.5% CTF113 5873.3 0.0244.2E−02 13.5% Control 0.02 0.0% Control 0.021 0.0% Table 27: Analyses ofseed yield and weight [expressed as 1000-seed weight in grams (g)] oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35S;SEQ ID NO: 675) when grown under high salinity irrigation conditions(80-100 mM NaCl) as compared to control plants. “Incr.” = increment withrespect to a control plant which has been transformed with an emptyvector. Ave. = Average calculated from several transgenic events. “Event#” = number of event (transgenic transformation).

TABLE 28 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved harvest index undernitrogen deficient growth conditions Harvest Index Gene Name Event #Average p-value % increment CTF113 5871.1 0.05 2.4E−02 67.8% CTF1135871.2 0.04 4.9E−02 42.8% Control 0.03 0.0% Table 28: Analyses ofharvest index of transgenic plants transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S; SEQ ID NO: 675) when grownunder high salinity irrigation conditions (80-100 mM NaCl) as comparedto control plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

TABLE 29 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved dry weight under highsalinity irrigationv growth conditions Dry Weight Gene Name Event #Average p-value % increment CTF113 5871.2 0.82 4.3E−03 23.7% CTF1135873.3 0.83 2.9E−03 26.0% Control 0.66 0.0% Table 29: Analyses of dryweight of transgenic plants transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S; SEQ ID NO: 675) when grownunder high salinity irrigation conditions (80-100 mM NaCl) as comparedto control plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

The genes presented in Tables 30-31, hereinbelow, have improved plantperformance and under regular growth conditions since they producedhigher seed yield, harvest index, seed weight (expressed as 1000-seedweight) and plant biomass [(as expressed as plant dry weight (DW)] whengrown under standard growth conditions, compared to control plants.

Tables 30-31 depict analyses of dry weight and seed yield (Table 30) andharvest index and seed weight (expressed as 1000-seed weight; Table 31)when grown under standard conditions (6 mM KNO₃, 1 mM KH₂PO₄, 1 mMMgSO₄, 2 mM CaCl₂ and microelements) in plants overexpressing thepolynucleotides of some embodiments of the invention under theregulation of a constitutive promoter (35S; SEQ ID NO:675). Evaluationof each gene was performed by testing the performance of several events.Some of the genes were evaluated in more than one tissue culture assayand the results obtained were repeated. Event with p-value <0.05 wasconsidered statistically significant.

TABLE 30 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved plant biomass (dryweight) and seed yield under standard conditions Gene Dry Weight (g)Gene Seed Yield (g) Name Event # Ave. P-Value % incr. Name Event # Ave.P-Value % incr. BDL103 7853.3 0.94 3.3E−02 37.0% BDL103 7853.1 0.301.3E−02 32.4% BDL103 8033.4 0.98 3.3E−02 42.3% BDL103 8033.4 0.302.6E−02 36.2% Control 0.69 0.0% Control 0.22 0.0% BDL103 8033.3 1.322.5E−02 31.8% BDL103 8033.3 0.71 1.2E−04 43.7% Control 1.00 0.0% Control0.49 0.0% Table 30: Analyses of plant biomass (dry weight) and seedyield of transgenic plants overexpressing the exogenous polynucleotidesof some embodiments of the invention (using the cloned or syntheticgenes listed in Table 10 above) under the regulation of a constitutivepromoter (35S; SEQ ID NO: 675) when grown under normal growth conditions(6 mM KNO₃, 1 mM KH₂PO₄, 1 mM MgSO₄, 2 mM CaCl₂ and microelements) ascompared to control plants. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

TABLE 31 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention under theregulation of the 35S promoter exhibit improved harvest index and seedweight under standard nitrogen conditions Gene Harvest Index Gene SeedWeight (g) Name Event# Ave. P-Value % incr. Name Event # Ave. P-Value %incr. BDL14 5762.1 0.31 1.2E−02 45.1% BDL103 7221.1 0.022 4.1E−03 22.3%BDL14 5763.2 0.27 3.5E−02 29.6% BDL103 7855.2 0.021 9.0E−03 18.3%Control 0.21 0.0% BDL103 8033.12 0.021 4.1E−02 19.5% BDL14 5762.1 0.533.9E−02 8.4% BDL103 8033.4 0.020 4.8E−02 13.3% Control 0.49 0.0% Control0.018 0.0% Table 31: Analyses of harvest index and seed weight oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35S;SEQ ID NO: 675) when grown under standard nitrogen conditions (6 mMKNO₃, 1 mM KH₂PO₄, 1 mM MgSO₄, 2 mM CaCl₂ and microelements) as comparedto control plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

Example 8 Evaluation of Transgenic Arabidopsis Plant Growth UnderAbiotic Stress as Well as Under Favorable Conditions in Greenhouse AssayGrown Until Bolting

This assay follows seed yield production, the biomass formation and therosette area growth of plants grown in the greenhouse at high salinityand regular growth conditions. Transgenic Arabidopsis seeds were sown inagar media supplemented with ½ MS medium and a selection agent(Kanamycin). The T₂ transgenic seedlings were then transplanted to 1.7trays filled with peat and perlite. The trays were irrigated with tapwater (provided from the pots' bottom). Half of the plants wereirrigated with a salt solution (50-150 mM NaCl and 5 mM CaCl₂) so as toinduce salinity stress (stress conditions). The other half of the plantswas irrigated with tap water (normal conditions). All plants were grownin the greenhouse until 90% of plants reach bolting (inflorescent startto emerge). Plant biomass (the above ground tissue) was weightedimmediately after harvesting the rosette (plant fresh weight [FW]).Following, plants were dried in an oven at 50° C. for 48 hours andweighted (plant dry weight [DW]).

Each construct was validated at its T₂ generation. Transgenic plantstransformed with a construct conformed by an empty vector carrying the35S promoter and the selectable marker was used as control.

The plants were analyzed for their overall size, growth rate, freshweight and dry matter. Transgenic plants performance was compared tocontrol plants grown in parallel under the same conditions.

The experiment was planned in nested randomized plot distribution. Foreach gene of the invention three to five independent transformationevents were analyzed from each construct.

Digital Imaging—

A laboratory image acquisition system, which consists of a digitalreflex camera (Canon EOS 300D) attached with a 55 mm focal length lens(Canon EF-S series), mounted on a reproduction device (Kaiser RS), whichincludes 4 light units (4×150 Watts light bulb) was used for capturingimages of plant samples.

The image capturing process was repeated every 2 days starting from day1 after transplanting till day 15. Same camera, placed in a custom madeiron mount, was used for capturing images of larger plants sawn in whitetubs in an environmental controlled greenhouse. During the captureprocess, the tubes were placed beneath the iron mount, while avoidingdirect sun light and casting of shadows.

An image analysis system was used, which consists of a personal desktopcomputer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ1.39 [Java based image processing program which was developed at theU.S. National Institutes of Health and freely available on the internetat Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/]. Imageswere captured in resolution of 10 Mega Pixels (3888×2592 pixels) andstored in a low compression JPEG (Joint Photographic Experts Groupstandard) format. Next, analyzed data was saved to text files andprocessed using the JMP statistical analysis software (SAS institute).

Leaf Analysis—

Using the digital analysis leaves data was calculated, including leafnumber, rosette area, rosette diameter, leaf blade area, plot coverageand leaf petiole area.

Vegetative growth rate: is the rate of growth of the plant as defined byformula VIII, IX, X and XI as described in Example 7 hereinabove.

Plant Fresh and Dry Weight—

On about day 40 from sowing, the plants were harvested and directlyweighted for the determination of the plant fresh weight (FW) and leftto dry at 50° C. in a drying chamber for about 48 hours before weightingto determine plant dry weight (DW).

Statistical Analyses—

To identify genes conferring significantly improved ABST, the resultsobtained from the transgenic plants were compared to those obtained fromcontrol plants. To identify outperforming genes and constructs, resultsfrom the independent transformation events tested are analyzedseparately. Data was analyzed using Student's t-test and results wereconsidered significant if the p value was less than 0.1. The JMPstatistics software package was used (Version 5.2.1, SAS Institute Inc.,Cary, N.C., USA).

Experimental Results:

The genes presented in Tables 32-36, hereinbelow, were found to increaseABST when grown under high salinity irrigation conditions, compared tocontrol plants. These genes produced larger plants with a largerphotosynthetic capacity when grown under limiting nitrogen conditions.

Tables 32-36 depict analyses of plant biomass and photosynthetic area(fresh weight, dry weight, rosette diameter, rosette area and plotcoverage) when grown under high salinity irrigation conditions (80-150mM NaCl) in plants overexpressing the polynucleotides of someembodiments of the invention under the regulation of a constitutivepromoter (At6669; SEQ ID NO:674). Evaluation of each gene was performedby testing the performance of several events. Some of the genes wereevaluated in more than one tissue culture assay and the results obtainedwere repeated. Event with p-value <0.05 was considered statisticallysignificant.

TABLE 32 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under high salinity conditions Fresh weight (g) Dry weight(g) Gene Gene Name Event # Ave. p-value % incr. Name Event # Ave.p-value % incr. LAB22 11064.6 0.57 3.0E−06 75.1% LAB22 11064.6 0.071.0E−03 86.8% LAB41 11552.1 0.40 1.8E−02 22.2% LAB41 11551.2 0.072.4E−02 113.2% Control 0.33 0.0% Control 0.03 0.0% LAB22 11062.1 0.571.5E−02 11.2% LAB21 11144.1 0.07 6.4E−03 22.8% LAB21 11144.1 0.639.3E−03 21.2% LAB34 11175.1 0.07 1.8E−02 35.7% LAB25 11341.2 0.643.4E−02 24.8% LAB33 11272.4 0.06 2.0E−02 15.8% LAB17 11533.1 0.588.1E−03 12.7% LAB33 11273.1 0.06 2.0E−02 15.8% LAB17 11533.6 0.661.5E−02 27.8% LAB25 11341.2 0.06 1.3E−02 18.1% LAB15 11641.1 0.563.9E−02 7.9% LAB17 11533.6 0.07 1.9E−03 32.3% Control 0.52 0.0% LAB2311571.2 0.08 1.2E−02 42.7% LAB40 11154.1 0.75 1.2E−02 20.8% LAB2311571.5 0.07 5.7E−03 24.0% LAB40 11154.4 0.86 8.2E−03 38.4% LAB1511642.2 0.07 3.8E−02 36.8% LAB40 11154.5 0.76 2.0E−02 23.2% Control 0.050.0% LAB24 11193.1 0.84 3.9E−03 35.4% LAB40 11151.1 0.09 1.4E−02 24.7%LAB49 11281.2 0.84 1.0E−03 35.4% LAB40 11154.5 0.08 4.2E−02 16.7% LAB311333.9 0.83 5.0E−03 33.3% LAB24 11193.1 0.09 3.9E−02 28.9% LAB1411471.1 0.92 2.9E−04 48.5% LAB24 11193.5 0.09 2.5E−03 31.6% LAB1411474.1 0.87 7.6E−04 40.4% LAB3 11333.9 0.08 4.6E−02 17.5% LAB14 11474.30.76 4.5E−02 22.2% LAB35 11461.2 0.08 3.1E−02 17.5% LAB51 11563.1 0.831.4E−03 34.3% LAB14 11471.1 0.10 9.4E−04 38.6% Control 0.62 0.0% LAB1411474.1 0.09 1.3E−02 24.6% LAB35 11462.3 0.77 1.7E−02 14.3% LAB5111561.2 0.10 5.7E−03 43.9% LAB35 11462.5 0.75 3.9E−02 11.5% LAB5111563.1 0.09 9.7E−03 26.3% LAB14 11472.1 0.88 7.8E−04 30.0% Control 0.070.0% Control 0.67 0.0% LAB35 11462.3 0.09 2.3E−02 22.4% LAB35 11462.50.09 3.3E−02 18.9% LAB14 11472.1 0.09 1.2E−02 30.2% Control 0.07 0.0%Table 32: Analyses of fresh weight and dry weight of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (6669) when grown underhigh salinity conditions as compared to control plants. “g” = grams.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 33 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under high salinity conditions Rosette diameter (cm)Rosette area (cm²) Gene Gene Name Event # Ave. p-value % incr. NameEvent # Ave. p-value % incr. LAB22 11064.6 2.28 3.4E−02 25.5% LAB2211064.6 1.88 2.6E−03 49.0% LAB41 11551.2 2.49 1.9E−02 37.3% LAB4111551.2 2.14 3.3E−02 70.0% LAB41 11552.1 2.10 9.9E−03 15.8% LAB4111552.1 1.50 4.1E−02 18.8% Control 1.81 0.0% Control 1.26 0.0% LAB2111144.1 2.69 2.4E−02 12.6% LAB21 11144.1 2.31 1.1E−02 27.4% LAB2511341.2 2.77 7.1E−03 16.2% LAB25 11341.2 2.44 7.1E−03 34.4% LAB1711531.6 2.67 9.1E−03 12.1% LAB17 11531.6 2.17 3.3E−02 19.7% LAB1511642.2 2.80 5.2E−03 17.4% LAB17 11533.6 2.65 8.5E−03 45.7% Control 2.390.0% LAB15 11642.2 2.45 6.2E−03 35.1% LAB40 11154.4 2.92 2.6E−03 24.3%Control 1.82 0.0% LAB40 11154.5 2.80 6.5E−03 19.2% LAB40 11151.1 2.421.6E−02 44.8% LAB24 11193.1 2.70 2.9E−02 15.3% LAB40 11154.4 2.822.2E−02 68.2% LAB24 11193.5 2.71 4.0E−02 15.7% LAB40 11154.5 2.363.6E−03 40.7% LAB49 11281.2 2.72 1.3E−02 16.0% LAB24 11193.1 2.173.9E−02 29.9% LAB5 11443.4 2.63 3.5E−02 12.2% LAB49 11281.2 2.42 2.6E−0344.8% LAB5 11444.1 3.02 1.0E−03 28.8% LAB3 11333.9 2.43 3.3E−02 45.4%LAB5 11444.5 2.86 3.5E−03 21.9% LAB5 11443.4 2.18 1.3E−02 30.2% LAB5111561.1 2.74 3.1E−02 17.0% LAB5 11444.1 2.78 8.6E−03 66.1% LAB51 11561.22.81 2.5E−02 19.8% LAB5 11444.5 2.52 2.4E−02 50.3% LAB51 11563.1 2.805.7E−03 19.3% LAB51 11561.1 2.27 6.6E−03 35.5% Control 2.34 0.0% LAB5111561.2 2.47 2.5E−03 47.6% LAB51 11563.1 2.37 2.1E−02 41.5% Control 1.670.0% Table 33: Analyses of rosette diameter and area of transgenicplants overexpressing the exogenous polynucleotides of some embodimentsof the invention (using the cloned or synthetic genes listed in Table 10above) under the regulation of a constitutive promoter (6669) when grownunder high salinity conditions as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 34 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under high salinity conditions Plot coverage (cm²) Leafnumber Gene Gene Name Event # Ave. p-value % incr. Name Event # Ave.p-value % incr. LAB22 11064.6 15.03 2.6E−03 49.0% LAB16 11033.2 8.135.0E−04 12.3% LAB41 11551.2 17.15 3.3E−02 70.0% LAB16 11034.1 7.942.3E−03 9.7% LAB41 11552.1 11.98 4.1E−02 18.8% LAB22 11062.1 7.633.2E−02 5.4% Control 10.09 0.0% LAB22 11064.6 8.00 1.3E−02 10.6% LAB2111144.1 18.51 1.1E−02 27.4% Control 7.23 0.0% LAB25 11341.2 19.537.1E−03 34.4% LAB5 11441.1 8.41 1.7E−02 13.1% LAB17 11531.6 17.403.3E−02 19.7% Control 7.44 0.0% LAB17 11533.6 21.17 8.5E−03 45.7% LAB1511642.2 19.62 6.2E−03 35.1% Control 14.53 0.0% LAB40 11154.1 16.793.3E−02 25.4% LAB40 11154.4 22.53 2.2E−02 68.2% LAB40 11154.5 18.843.6E−03 40.7% LAB24 11193.1 17.40 3.9E−02 29.9% LAB49 11281.2 19.402.6E−03 44.8% LAB3 11333.9 19.48 3.3E−02 45.4% LAB5 11443.4 17.441.3E−02 30.2% LAB5 11444.1 22.24 8.6E−03 66.1% LAB5 11444.5 20.132.4E−02 50.3% LAB51 11561.1 18.15 6.6E−03 35.5% LAB51 11561.2 19.772.5E−03 47.6% LAB51 11563.1 18.95 2.1E−02 41.5% Control 13.39 0.0% Table34: Analyses of plot coverage and leaf number of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 32 above)under the regulation of a constitutive promoter (6669) when grown underhigh salinity conditions as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 35 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass and growth rate under high salinity conditions RGR ofrosette area RGR of rosette diameter (Regression coefficient)(Regression coefficient) Gene Gene Name Event # Average p-value % incr.Name Event # Average p-value % incr. LAB11 11024.3 0.24 2.9E−02 46.4%LAB11 11024.3 0.23 2.1E−02 32.4% LAB16 11033.2 0.29 1.6E−03 75.5% LAB1611033.2 0.25 2.4E−03 44.8% LAB16 11034.1 0.33 4.9E−05 101.2% LAB1611034.1 0.28 8.6E−05 58.1% LAB22 11062.1 0.26 5.9E−03 59.3% LAB1611034.4 0.24 7.6E−03 39.2% LAB22 11062.3 0.25 2.5E−02 51.6% LAB2211062.1 0.24 5.1E−03 37.9% LAB22 11064.6 0.25 6.5E−03 55.1% LAB2211064.6 0.24 3.6E−03 37.4% LAB41 11551.2 0.29 4.4E−04 76.7% LAB4111551.2 0.26 4.5E−04 47.8% Control 0.16 0.0% Control 0.17 0.0% LAB2111144.1 0.30 3.9E−02 27.0% LAB25 11341.2 0.28 4.7E−02 15.9% LAB2511341.2 0.32 1.4E−02 34.6% LAB13 Control 0.24 0.0% LAB17 11533.6 0.342.5E−03 43.8% LAB3 11333.9 0.30 3.9E−02 25.6% LAB17 11534.1 0.31 4.4E−0229.0% Control 0.24 0.0% LAB23 11571.2 0.34 1.4E−02 40.4% LAB23 11571.50.32 1.1E−02 34.6% LAB15 11642.2 0.32 1.2E−02 33.1% Control 0.24 0.0%LAB40 11151.1 0.31 2.5E−02 45.7% LAB40 11154.4 0.36 2.7E−03 65.8% LAB4911281.2 0.31 4.4E−02 41.6% LAB49 11281.4 0.35 6.3E−03 62.6% LAB3 11333.90.31 3.3E−02 45.8% LAB5 11444.1 0.36 3.5E−03 65.6% LAB5 11444.5 0.321.8E−02 50.1% LAB51 11561.2 0.32 2.6E−02 47.4% LAB51 11563.1 0.313.9E−02 42.9% Control 0.22 0.0% Table 35: Analyses of relative growthrate (RGR) of rosette area and diameter of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (6669) when grown underhigh salinity conditions as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

TABLE 36 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass and growth rate under high salinity conditions RGR of plotcoverage RGR of plot coverage Gene Gene Name Event # Ave. p-value %incr. Name Event # Ave. p-value % incr. LAB11 11024.3 1.90 2.9E−02 46.4%LAB40 11151.1 2.80 6.3E−03 62.6% LAB16 11033.2 2.28 1.6E−03 75.5% LAB4011154.4 2.51 3.3E−02 45.8% LAB16 11034.1 2.61 4.9E−05 101.2% LAB4911281.2 2.85 3.5E−03 65.6% LAB22 11062.1 2.07 5.9E−03 59.3% LAB4911281.4 2.59 1.8E−02 50.1% LAB22 11062.3 1.97 2.5E−02 51.6% LAB3 11333.92.54 2.6E−02 47.4% LAB22 11064.6 2.02 6.5E−03 55.1% LAB5 11444.1 2.463.9E−02 42.9% LAB41 11551.2 2.30 4.4E−04 76.7% LAB5 11444.5 1.90 2.9E−0246.4% Control 2.43 3.9E−02 27.0% LAB51 11561.2 2.28 1.6E−03 75.5% LAB2111144.1 2.58 1.4E−02 34.6% LAB51 11563.1 2.61 4.9E−05 101.2% LAB2511341.2 2.76 2.5E−03 43.8% Control 2.07 5.9E−03 59.3% LAB17 11533.6 2.474.4E−02 29.0% LAB40 11151.1 2.80 6.3E−03 62.6% LAB17 11534.1 2.691.4E−02 40.4% LAB40 11154.4 2.51 3.3E−02 45.8% LAB23 11571.2 2.581.1E−02 34.6% LAB49 11281.2 2.85 3.5E−03 65.6% LAB23 11571.5 2.551.2E−02 33.1% LAB49 11281.4 2.59 1.8E−02 50.1% LAB15 11642.2 2.862.7E−03 65.8% LAB3 11333.9 2.54 2.6E−02 47.4% Control 2.44 4.4E−02 41.6%LAB5 11444.1 2.46 3.9E−02 42.9% LAB5 11444.5 1.90 2.9E−02 46.4% LAB5111561.2 2.28 1.6E−03 75.5% LAB51 11563.1 2.61 4.9E−05 101.2% Control2.07 5.9E−03 59.3% Table 36: Analyses of relative growth rate (RGR) ofplot coverage of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (6669) when grown under high salinity conditionsas compared to control plants. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

Tables 37-41 depict analyses of plant biomass, growth rate andphotosynthetic area (fresh weight, dry weight, rosette diameter, rosettearea and plot coverage) when grown under normal conditions in plantsoverexpressing the polynucleotides of some embodiments of the inventionunder the regulation of a constitutive promoter (At6669; SEQ ID NO:674).Evaluation of each gene was performed by testing the performance ofseveral events. Some of the genes were evaluated in more than one tissueculture assay and the results obtained were repeated. Event with p-value<0.05 was considered statistically significant.

TABLE 37 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under normal conditions Fresh weight (g) Dry weight (g)Gene Gene Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. LAB16 11033.2 1.13 2.4E−02 125.3% LAB16 11033.2 0.11 0.0E+00111.5% LAB22 11064.6 1.19 0.0E+00 139.1% LAB22 11062.1 0.09 4.0E−0672.2% LAB11 Control 0.50 0.0% LAB22 11064.6 0.11 0.0E+00 114.0% LAB3811434.3 2.59 4.5E−02 28.7% Control 0.05 0.0% Control 2.02 0.0% LAB4011151.1 0.16 6.7E−03 18.8% LAB40 11151.1 1.81 2.9E−02 16.9% LAB4011154.5 0.15 1.7E−02 13.1% LAB40 11154.5 1.81 2.9E−02 16.9% LAB3911182.1 0.17 5.6E−03 27.2% LAB39 11182.1 1.98 1.4E−03 27.4% LAB2411193.1 0.16 6.3E−03 16.9% LAB24 11193.1 1.84 1.2E−02 19.0% LAB4911281.6 0.18 1.8E−04 36.6% LAB49 11281.6 2.19 2.6E−05 41.1% LAB5 11444.10.16 5.2E−03 23.0% LAB3 11331.1 1.74 1.4E−02 12.1% LAB35 11461.2 0.191.4E−02 46.0% LAB5 11444.1 2.07 8.1E−03 33.5% LAB14 11471.1 0.15 1.8E−0213.6% LAB35 11461.2 2.30 1.0E−05 48.4% LAB51 11561.5 0.15 2.3E−02 15.5%LAB35 11462.5 1.91 1.2E−03 23.0% Control 0.13 0.0% LAB14 11474.1 1.803.3E−02 16.1% Control 1.55 0.0% LAB49 11281.6 1.84 3.8E−02 9.9% Control1.68 0.0% Table 37: Analyses of fresh weight and dry weight oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter(At6669; SEQ ID NO: 674) when grown under noraml conditions as comparedto control plants, “g” = grams. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

TABLE 38 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass normal conditions Rosette diameter (cm) Rosette area (cm²)Gene Gene Name Event # Ave. p-value % Incr. Name Event # Ave. p-value %Incr. LAB16 11033.2 3.00 3.5E−02 73.6% LAB16 11033.2 2.91 9.5E−03 150.9%LAB22 11064.6 2.84 0.0E+00 64.4% LAB22 11064.6 2.85 0.0E+00 145.9% LAB4111551.4 1.95 3.7E−02 12.8% Control 1.16 0.0% Control 1.73 0.0% LAB3811434.3 4.48 4.1E−02 46.7% LAB40 11151.1 3.31 6.5E−03 17.4% Control 3.050.0% LAB40 11154.1 3.82 2.3E−02 35.7% LAB40 11151.1 3.19 2.4E−04 42.6%LAB39 11182.1 3.30 1.0E−03 17.2% LAB39 11182.1 3.05 5.8E−03 36.0% LAB2411192.1 3.23 2.4E−03 14.7% LAB24 11192.1 2.90 1.9E−03 29.5% LAB2411193.1 3.21 2.8E−03 13.9% LAB24 11193.1 2.77 1.2E−02 23.6% LAB4911281.4 3.33 2.0E−03 18.1% LAB49 11281.4 2.91 1.7E−03 29.7% LAB4911281.6 3.57 1.5E−04 26.6% LAB49 11281.6 3.26 1.8E−04 45.4% LAB3 11333.13.22 2.4E−03 14.3% LAB3 11333.1 2.76 2.3E−02 23.1% LAB3 11333.9 3.245.2E−03 15.1% LAB3 11333.9 2.92 4.1E−02 30.4% LAB5 11444.1 3.42 3.6E−0421.3% LAB5 11443.3 3.19 1.8E−02 42.4% LAB35 11461.2 3.81 1.8E−02 35.3%LAB5 11444.1 3.24 2.5E−04 44.7% LAB51 11561.5 3.30 1.3E−03 17.0% LAB3511461.2 4.25 1.8E−02 89.8% Control 2.82 0.0% LAB51 11561.5 2.77 9.7E−0323.6% Control 2.24 0.0% Table 38: Analyses of rosette diameter and areaof transgenic plants overexpressing the exogenous polynucleotides ofsome embodiments of the invention (using the cloned or synthetic geneslisted in Table 10 above) under the regulation of a constitutivepromoter (At6669; SEQ ID NO: 674) when grown under normal conditions ascompared to control plants. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

TABLE 39 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under normal conditions Plot coverage (cm²) Leaf numberGene Gene Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. LAB16 11033.2 23.25 9.5E−03 150.9% LAB16 11032.5 8.44 4.1E−0210.9% LAB22 11064.6 22.79 0.0E+00 145.9% LAB22 11064.6 8.69 9.0E−0414.2% Control 9.27 0.0% Control 7.61 0.0% LAB38 11434.3 35.81 4.1E−0246.7% LAB39 11182.1 8.25 3.2E−02 8.5% Control 24.41 0.0% LAB49 11283.68.31 1.1E−02 9.3% LAB40 11151.1 25.56 2.4E−04 42.6% LAB35 11461.2 9.001.3E−03 18.4% LAB39 11182.1 24.38 5.8E−03 36.0% Control 7.60 0.0% LAB2411192.1 23.21 1.9E−03 29.5% LAB24 11193.1 22.15 1.2E−02 23.6% LAB4911281.4 23.25 1.7E−03 29.7% LAB49 11281.6 26.05 1.8E−04 45.4% LAB311333.1 22.06 2.3E−02 23.1% LAB3 11333.9 23.38 4.1E−02 30.4% LAB511443.3 25.53 1.8E−02 42.4% LAB5 11444.1 25.93 2.5E−04 44.7% LAB3511461.2 34.01 1.8E−02 89.8% LAB51 11561.5 22.16 9.7E−03 23.6% Control17.92 0.0% Table 39: Analyses of plot coverage and leaf number oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter(At6669; SEQ ID NO: 674) when grown under normal conditions as comparedto control plants. “Incr.” = increment with respect to a control plantwhich has been transformed with an empty vector. Ave. = Averagecalculated from several transgenic events. “Event #” = number of event(transgenic transformation).

TABLE 40 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass and growth rate under normal conditions RGR of rosettearea RGR of rosette diameter (Regression coefficient) (Regressioncoefficient) Gene Gene Name Event # Ave. p-value % incr. Name Event #Ave. p-value % incr. LAB16 11032.5 0.33 9.2E−04 121.8% LAB16 11032.50.27 1.7E−03 65.7% LAB16 11033.2 0.39 0.0E+00 163.5% LAB16 11033.2 0.320.0E+00 97.3% LAB22 11062.1 0.30 4.3E−05 101.9% LAB22 11062.1 0.293.0E−06 77.5% LAB22 11063.4 0.29 7.0E−03 93.7% LAB22 11063.4 0.258.5E−03 55.1% LAB22 11064.1 0.21 4.9E−02 43.5% LAB22 11064.1 0.213.2E−02 29.1% LAB22 11064.6 0.39 0.0E+00 161.4% LAB22 11064.6 0.310.0E+00 92.7% Control 0.15 0.0% LAB41 11551.4 0.20 4.0E−02 22.1% LAB1711534.1 0.66 4.6E−02 60.4% Control 0.16 0.0% Control 0.41 0.0% LAB4011154.1 0.39 5.1E−03 31.3% LAB40 11151.1 0.42 2.6E−02 42.0% LAB4911281.6 0.38 9.4E−03 29.2% LAB40 11154.1 0.51 7.6E−04 75.6% LAB3511461.2 0.39 6.0E−03 31.7% LAB39 11182.1 0.41 3.6E−02 38.3% Control 0.290.0% LAB49 11281.6 0.43 2.0E−02 45.4% LAB5 11443.3 0.42 2.1E−02 43.5%LAB5 11444.1 0.42 2.2E−02 44.5% LAB5 11444.5 0.45 1.9E−02 53.3% LAB3511461.2 0.56 1.6E−04 89.8% Control 0.29 0.0% Table 40: Analyses ofrelative growth rate (RGR) of rosette area and diameter of transgenicplants overexpressing the exogenous polynucleotides of some embodimentsof the invention (using the cloned or synthetic genes listed in Table 10above) under the regulation of a constitutive promoter (At6669; SEQ IDNO: 674) when grown under normal conditions as compared to controlplants. “Incr.” = increment with respect to a control plant which hasbeen transformed with an empty vector. Ave. = Average calculated fromseveral transgenic events. “Event #” = number of event (transgenictransformation).

TABLE 41 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass and growth rate under normal conditions RGR of plotcoverage RGR of plot coverage Gene Gene Name Event # Ave. p-value %incr. Name Event # Ave. p-value % incr. LAB16 11032.5 2.64 9.2E−04121.8% LAB40 11151.1 3.33 2.6E−02 42.0% LAB16 11033.2 3.14 0.0E+00163.5% LAB40 11154.1 4.12 7.6E−04 75.6% LAB22 11062.1 2.40 4.3E−05101.9% LAB39 11182.1 3.24 3.6E−02 38.3% LAB22 11063.4 2.30 7.0E−03 93.7%LAB49 11281.6 3.41 2.0E−02 45.4% LAB22 11064.1 1.71 4.9E−02 43.5% LAB511443.3 3.36 2.1E−02 43.5% LAB22 11064.6 3.11 0.0E+00 161.4% LAB511444.1 3.39 2.2E−02 44.5% Control 1.19 0.0% LAB5 11444.5 3.59 1.9E−0253.3% LAB17 11534.1 5.31 4.6E−02 60.4% LAB35 11461.2 4.45 1.6E−04 89.8%Control 3.31 0.0% Control 2.34 0.0% Table 41: Analyses of relativegrowth rate (RGR) of plot coverage of transgenic plants overexpressingthe exogenous polynucleotides of some embodiments of the invention(using the cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (At6669; SEQ ID NO: 674) whengrown under normal conditions as compared to control plants. “Incr.” =increment with respect to a control plant which has been transformedwith an empty vector. Ave. = Average calculated from several transgenicevents. “Event #” = number of event (transgenic transformation).

Tables 42-46 depict analyses of plant biomass and photosynthetic area(fresh weight, dry weight, rosette diameter, rosette area and plotcoverage) when grown under normal conditions in plants overexpressingthe polynucleotides of some embodiments of the invention under theregulation of a constitutive promoter (35S; SEQ ID NO:675). Evaluationof each gene was performed by testing the performance of several events.Some of the genes were evaluated in more than one tissue culture assayand the results obtained were repeated. Event with p-value <0.05 wasconsidered statistically significant.

TABLE 42 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under normal conditions Fresh weight (g) Dry weight (g)Gene Gene Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. BDL210 10834.3 1.83 4.6E−02 17.1% BDL210 10831.3 0.16 3.9E−0216.4% Control 1.56 0.0% BDL210 10833.1 0.19 4.5E−02 32.0% CTF226 10985.21.90 4.3E−02 10.7% BDL210 10834.3 0.17 8.8E−03 23.1% Control 1.72 0.0%Control 0.14 0.0% Table 42: Analyses of fresh and dry weight oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35S;SEQ ID NO: 675) when grown under noraml conditions as compared tocontrol plants. “g” = grams. “Incr.” = increment with respect to acontrol plant which has been transformed with an empty vector. Ave. =Average calculated from several transgenic events. “Event #” = number ofevent (transgenic transformation).

TABLE 43 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass normal conditions Rosette diameter (cm) Rosette area (cm²)Gene Gene Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. BDL210 10831.3 3.65 2.7E−02 6.5% BDL210 10831.3 4.21 1.5E−02 16.9%Control 3.43 0.0% BDL210 10834.2 4.21 4.8E−02 17.0% Control 3.60 0.0%Table 43: Analyses of rosette diameter and area of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S; SEQ ID NO: 675)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 44 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under normal conditions Plot coverage (cm²) Leaf numberGene Gene Name Event # Ave. p-value % incr. Name Event # Ave. p-value %incr. BDL210 10831.3 33.67 1.5E−02 16.9% BDL210 10833.1 9.31 6.4E−036.2% BDL210 10834.2 33.70 4.8E−02 17.0% Control 8.77 0.0% 28.80 0.0%Table 44: Analyses of plot coverage and leaf number of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S; SEQ ID NO: 675)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 45 Transgenic plants exogenously expressing the polynucleotides ofsome embodiments of the invention exhibit improved plant biomass undernormal conditions RGR of rosette area RGR of rosette diameter Gene GeneName Event # Ave. p-value % incr. Name Event # Ave. p-value % incr.BDL210 10833.1 0.81 2.4E−03 66.5% BDL210 10833.1 0.46 3.8E−02 21.9%Control 0.49 0.0% Control 0.38 0.0% Table 45: Analyses of relativegrowth rate (RGR) of rosette area and diameter of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S; SEQ ID NO: 675)when grown under normal conditions as compared to control plants.“Incr.” = increment with respect to a control plant which has beentransformed with an empty vector. Ave. = Average calculated from severaltransgenic events. “Event #” = number of event (transgenictransformation).

TABLE 46 Transgenic Arabidopsis plants exogenously expressing thepolynucleotides of some embodiments of the invention exhibit improvedplant biomass under normal conditions RGR of plot coverage Gene NameEvent # Ave. p-value % incr. BDL210 10833.1 6.50 2.4E−03 66.5% Control3.90 0.0% Table 46: Analyses of relative growth rate (RGR) of plotcoverage of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S SEQ ID NO: 675) when grown under normalconditions as compared to control plants. “Incr.” = increment withrespect to a control plant which has been transformed with an emptyvector. Ave. = Average calculated from several transgenic events. “Event#” = number of event (transgenic transformation).

Example 9 Improved Transgenic Plant Performance Under Normal Conditions

To analyze whether the transgenic plants has performed better, plantswere grown in pots with an adequate amount of nutrient and water. Theplants were analyzed for their overall size, growth rate, time toinflorescence emergence (bolting) and flowering, seed yield, oil contentof seed, weight of 1,000 seeds, dry matter and harvest index (HI-seedyield/dry matter). Transgenic plants performance was compared to controlplants grown in parallel under the same conditions. Mock-transgenicplants expressing the uidA reporter gene (GUS-Intron) under the samepromoter were used as control.

Parameters were measured as described in Examples 6, 7 and 8 above.

Statistical Analyses—

To identify genes conferring significantly improved plant performance,the results obtained from the transgenic plants were compared to thoseobtained from control plants. Plant growth rate, plant area, time tobolt, time to flower, weight of 1,000 seeds, seed yield, total yield,oil yield, oil percent in seeds, dry matter, harvest index, rosette areaand growth rate data were analyzed using one-way ANOVA. To identifyoutperforming genes and constructs, results from mix of transformationevents or independent events tested were analyzed. The Least MeanSquares were calculated for each experiment. For gene versus controlanalysis T-test was applied, using significance of p<0.05. The JMPstatistics software package was used (Version 5.2.1, SAS Institute Inc.,Cary, N.C., USA).

Experimental Results

The polynucleotide sequences of the invention were assayed for a numberof commercially desired traits.

Tables 47-57 depict analyses of seed yield (Table 47), oil yield (Table48), dry matter (Table 49), harvest index (HI) (Tables 50 and 51),growth rate (Table 52), rosette area (Table 53), oil % in seed (Table54), weight of 1000 seeds (Tables 55 and 56) and total yield (Table 57)in plants overexpressing the polynucleotides of some embodiments of theinvention under the regulation of a constitutive (35S; SEQ ID NO:675) ora seed specific (napin; SEQ ID NO:676) promoter. Each Table representsan independent experiment, using at least 5 independent events per gene.Genes not connected by same letter as the control (A, B) aresignificantly different (p<0.05) from the control.

TABLE 47 Genes showing improved plant performance Arabidopsis: Seedyield Seed yield per plant (g) Under Significance % regula- Least(t-Test compare improve- Gene Id tion of Mean Sq to control) ment BDL1135S 0.420 A 4.2 BDL17 35S 0.426 A 5.8 CONTROL 35S 0.403 A 0.0 (GUSIntron) BDL12 35S 0.319 B 9.7 BDL14 35S 0.378 A 30.3 CONTROL 35S 0.290 B0.0 (GUS Intron) Table 47: Analyses of seed yield per plant oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35Spromoter; SEQ ID NO: 675) when grown under normal conditions as comparedto control plants. “Least Mean Sq” = Least Mean Square. “% improvement”relates to improvement of transgenic plant seed yield as compared tocontrol plants that have been transformed with a vector comprising GUSintron under the transcriptional regulation of the same promoter.

TABLE 48 Genes showing improved plant performance Arabidopsis: Oil yieldOil yield per plant (gr) Under Significance % regula- Least (t-Testcompare improve- Gene Id tion of Mean Sq to control) ment BDL11 35S 0.12A 7.0 BDL17 35S 0.12 A 6.5 CONTROL (GUS 35S 0.12 A 0.0 Intron) BDL12 35S0.100 B 14.2 BDL14 35S 0.114 A 31.1 CONTROL (GUS 35S 0.087 B 0.0 Intron)Table 48: Analyses of oil yield per plant of transgenic plantsoverexpressing the exogenous polynucleotides of some embodiments of theinvention (using the cloned or synthetic genes listed in Table 10 above)under the regulation of a constitutive promoter (35S promoter; SEQ IDNO: 675) when grown under normal conditions as compared to controlplants. “Least Mean Sq” = Least Mean Square. “% improvement” relates toimprovement of transgenic plant oil yiel as compared to control plantsthat have been transformed with a vector comprising GUS intron under thetranscriptional regulation of the same promoter.

TABLE 49 Genes showing improved plant performance Arabidopsis: Drymatter Dry matter per plant (gr) Under Significance % regula- Least(t-Test compare improve- Gene Id tion of Mean Sq to control) ment BDL1435S 1.0444 A 9.7 CONTROL (GUS 35S 0.9523 A 0.0 Intron) BDL11 35S 1.3638A 1.2 CONTROL (GUS 35S 1.3474 A 0.0 Intron) Table 49. Analyses of drymatter per plant of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S promoter; SEQ ID NO: 675) when grown undernormal conditions as compared to control plants. “Least Mean Sq” = LeastMean Square. “% improvement” relates to improvement of transgenic plantdry matter as compared to control plants that have been transformed witha vector comprising GUS intron under the transcriptional regulation ofthe same promoter.

TABLE 50 Genes showing improved plant performance Arabidopsis: harvestindex (HI) HI Under Significance % regula- Least (t-Test compareimprove- Gene Id tion of Mean Sq to control) ment BDL11 35S 0.3063 B 2.0BDL17 35S 0.3526 A 17.5 CONTROL (GUS 35S 0.3002 B 0.0 Intron) Table 50.Analyses of harvest index of transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S promoter; SEQ ID NO: 675)when grown under normal conditions as compared to control plants. “LeastMean Sq” = Least Mean Square. “% improvement” relates to improvement oftransgenic harvest index as compared to control plants that have beentransformed with a vector comprising GUS intron under thetranscriptional regulation of the same promoter.

TABLE 51 Genes showing improved plant performance Arabidopsis: Harvestindex Harvest index Under Significance % regula- (t-Test compareimprove- Gene Id tion of Mean to control) ment BDL103 35S 0.341 A 16.8CONTROL (GUS 35S 0.292 B 0 Intron) Table 51. Analyses of harvest indexof transgenic plants overexpressing the exogenous polynucleotides ofsome embodiments of the invention (using the cloned or synthetic geneslisted in Table 10 above) under the regulation of a constitutivepromoter (35S promoter; SEQ ID NO: 675) when grown under normalconditions as compared to control plants. “Least Mean Sq” = Least MeanSquare. “% improvement” relates to improvement of transgenic plantharvest index as compared to control plants that have been transformedwith a vector comprising GUS intron under the transcriptional regulationof the same promoter.

TABLE 52 Genes showing improved plant performance Arabidopsis: Growthrate Growth rate (cm²/day) Under Significance % regula- Least (t-Testcompare improve- Gene Id tion of Mean Sq to control) ment BDL14 35S 2.48A 6.4 CONTROL (GUS 35S 2.33 A 0.0 Intron) BDL11 35S 1.80 A 15.4 CONTROL(GUS 35S 1.56 A 0.0 Intron) BDL12 35S 1.58 B 2.0 BDL14 35S 1.95 A 26.3CONTROL (GUS 35S 1.55 B 0.0 Intron) Table 52. Analyses of growth rate oftransgenic plants overexpressing the exogenous polynucleotides of someembodiments of the invention (using the cloned or synthetic genes listedin Table 10 above) under the regulation of a constitutive promoter (35Spromoter; SEQ ID NO: 675) when grown under normal conditions as comparedto control plants. “Least Mean Sq” = Least Mean Square. “% improvement”relates to improvement of transgenic plant growth rate as compared tocontrol plants that have been transformed with a vector comprising GUSintron under the transcriptional regulation of the same promoter.

TABLE 53 Genes showing improved plant performance Arabidopsis: Rossetearea Rosette area (cm²) Under Significance % regula- Least (t-Testcompare improve- Gene Id tion of Mean Sq to control) ment BDL14 35S11.83 A 9.2 CONTROL (GUS 35S 10.83 B 0.0 Intron) BDL11 35S 14.09 A 13.2CONTROL (GUS 35S 12.44 A 0.0 Intron) BDL12 35S 7.92 B −2.5 BDL14 35S9.96 A 22.7 CONTROL (GUS 35S 8.12 B 0.0 Intron) Table 53: Analyses ofrosette area of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S promoter; SEQ ID NO: 675) when grown undernormal conditions as compared to control plants. “Least Mean Sq” = LeastMean Square. “% improvement” relates to improvement of transgenic plantrosette area as compared to control plants that have been transformedwith a vector comprising GUS intron under the transcriptional regulationof the same promoter. It should be noted that an increase in rosettearea means better soil coverage and reduced water loss from soil.Decrease in rosette area means more plants could be put per areaincreasing yield.

TABLE 54 Genes showing improved plant performance Arabidopsis: oil % inseed Oil % in seed Under Significance % regula- Least (t-Test compareimprove- Gene Id tion of Mean Sq to control) ment BDL14 35S 31.31 A 3.1CONTROL (GUS 35S 30.355 A 0.0 Intron) BDL11 35S 29.216 A 1.5 BDL17 35S28.904 A 0.4 CONTROL (GUS 35S 28.78 A 0 Intron) BDL12 35S 31.30 A 3.7BDL14 35S 30.27 A 0.3 CONTROL (GUS 35S 30.19 A 0.0 Intron) Table 54.Analyses of oil percent in seed of transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a constitutive promoter (35S promoter; SEQ ID NO: 675)when grown under normal conditions as compared to control plants. “LeastMean Sq” = Least Mean Square. “% improvement” relates to improvement oftransgenic plant oil percent in seed as compared to control plants thathave been transformed with a vector comprising GUS intron under thetranscriptional regulation of the same promoter.

TABLE 55 Genes showing improved plant performance Arabidopsis: weight of1,000 seeds Weight of 1000 seeds (gr) Under Significance % regula- Least(t-Test compare improve- Gene Id tion of Mean Sq to control) ment BDL1435S 0.019 B 6.1 CONTROL (GUS 35S 0.018 B 0.0 Intron) BDL11 35S 0.0235 A15.7 CONTROL (GUS 35S 0.0203 B 0 Intron) BDL12 35S 0.0234 A 0.1 CONTROL(GUS 35S 0.0234 A 0.0 Intron) Table 55. Analyses of weight of 1,000seeds of transgenic plants overexpressing the exogenous polynucleotidesof some embodiments of the invention (using the cloned or syntheticgenes listed in Table 10 above) under the regulation of a constitutivepromoter (35S promoter; SEQ ID NO: 675) when grown under normalconditions as compared to control plants. “Least Mean Sq” = Least MeanSquare. “% improvement” relates to improvement of transgenic plantweight of 1,000 seeds as compared to control plants that have beentransformed with a vector comprising GUS intron under thetranscriptional regulation of the same promoter.

TABLE 56 Genes showing improved plant performance Arabidopsis: weight of1,000 seeds Weight of 1000 seeds (gr) Under Significance % regula- Least(t-Test compare improve- Gene Id tion of Mean Sq to control) ment BDL14Napin 0.0227 A 2.3 CONTROL (GUS Napin 0.0222 A 0.0 Intron) BDL12 Napin0.0206 A 0.2 CONTROL (GUS Napin 0.0205 A 0.0 Intron) Table 56. Analysesof weight of 1,000 seeds of transgenic plants overexpressing theexogenous polynucleotides of some embodiments of the invention (usingthe cloned or synthetic genes listed in Table 10 above) under theregulation of a seed specific napin promoter (SEQ ID NO: 675) when grownunder normal conditions as compared to control plants. “Least Mean Sq” =Least Mean Square. “% improvement” relates to improvement of transgenicplant weight of 1,000 seeds as compared to control plants that have beentransformed with a vector comprising GUS intron under thetranscriptional regulation of the same promoter.

TABLE 57 Genes showing improved plant performance Arabidopsis: totalyield total yield (gr/plant) Under Significance % regula- (t-Testcompare improve- Gene Id tion of Mean to control) ment BDL103 35S 0.305A 10.1 CONTROL (GUS 35S 0.277 B 0 Intron) Table 57. Analyses of totalyield per plant of transgenic plants overexpressing the exogenouspolynucleotides of some embodiments of the invention (using the clonedor synthetic genes listed in Table 10 above) under the regulation of aconstitutive promoter (35S promoter; SEQ ID NO: 675) when grown undernormal conditions as compared to control plants. “Least Mean Sq” = LeastMean Square. “% improvement” relates to improvement of transgenic planttotal yield as compared to control plants that have been transformedwith a vector comprising GUS intron under the transcriptional regulationof the same promoter.

Example 10 Transgenic Arabidosis which Exogenously Express BDL103Exhibit Increased Commercially Desired Traits in a Tissue Culture Assay

Nitrogen Use Efficiency—

Tissue culture assays were performed as described in Example 6hereinabove for determining plant performance under normal (i.e., 15 mMnitrogen) or nitrogen deficiency (i.e., 0.75 mM nitrogen) conditions.

Abiotic Stress Tolerance—

To determine whether the transgenic plants exhibit increased toleranceto abiotic stress such as drought, an osmotic stress was induced byadding sorbitol or polyethylene glycol (PEG 8000) to the culturingmedium. Control and transgenic plants were germinated and grown inplant-agar plates for 10 days, after which they were transferred toplates containing either 1.5% PEG8000 or 500 mM of sorbitol. Plants weregrown under the osmotic stress conditions or the normal conditions forabout additional 10 days, during which various parameters which indicateplant characteristics were measured. The measured parameters [e.g.,plant weight (fresh and dry), yield, growth rate] were compared betweenthe control and transgenic plants.

Tables 58-60 depict analyses of root coverage, root length, growth rateof root coverage, growth rate of root length and biomass in plantsoverexpressing the BDL103-short (SEQ ID NO:671) and BDL103-long (SEQ IDNO:670) polynucleotides under the regulation of a constitutive (35S; SEQID NO:675) when grown under normal conditions (Table 58), under nitrogenlimiting conditions (Table 59), or under osmotic stress (15% PEG). EachTable includes data of several transformation events per gene. Resultswere considered significant if p-value was lower than 0.1 when comparedto control plants (which were transformed with a vector containing GUSreporter gene).

TABLE 58 Improved growth rate, root coverage, root length and biomass intransgenic Arabidopsis plants exogenously expressing BDL103 under normalconditions BDL103 Long or Short/Event No. Long/ Long/ Long/ Long/ Long/Short/ Short/ Short/ Short/ Short/ 3054 3055 3056 3057 3058 3060 30613062 3063 3064 Roots P 0.10 Coverage A 1.21 (time point 1) Roots P 0.080.25 Coverage A 1.36 1.12 (time point 6) Roots P 0.07 Coverage A 1.23(time point 9) Roots Length P 0.03 (time point 1) A 1.19 Roots Length P0.05 0.22 (time point 6) A 1.17 1.10 Roots Length P 0.01 (time point 9)A 1.15 GR (growth P 0.07 0.46 rate) of Roots A 1.47 1.10 Coverage (timepoint 6) GR of Roots P 0.04 Length (time A 1.31 point 6) RGR of P 0.060.70 0.13 0.02 0.10 Roots A 1.56 1.16 1.31 1.49 1.28 Coverage (timepoint 6) RGR of P 0.23 Roots A 1.87 Coverage (time point 9) RGR of P0.08 0.74 0.21 0.11 Roots Length A 1.37 1.11 1.16 1.13 (time point 6)RGR of P 0.23 0.41 Roots Length A 1.62 1.12 (time point 9) DW [gr] P0.56 (time point 1) A 1.13 DW [gr] P 0.56 (time point 6) A 1.13 DW [gr]P 0.56 (time point 9) A 1.13 Table 58. Analysis of growth parameters intissue culture conditions of transgenic plants overexpressingBDL103-Short polynucleotide (SEQ ID NO: 671) or BDL103-Longpolynucleotide (SEQ ID NO: 670) under the regulation of a constitutivepromoter (35S; SEQ ID NO: 675) when grown under normal conditions (15 mMnitrogen). Each event number refers to an independent transformationevent in a plant (i.e., generation of a transgenic plant expressing thepolynucleotide of choice). “A” = average; “P” = p-value; “GR” = growthrate; “RGR” = relative growth rate; “DW” = dry weight; “gr” = grams;Root coverage is presented in cm²; root length is presented in cm; GR ofroot length is presented in cm/day; RGR of root length is presented incm/day; RGR of root coverage is presented in cm²/day. The various timepoints indicate days from beginning of experiment in which parameterswere measured.

TABLE 59 Improved growth rate, root coverage, root length and biomass intransgenic plants exogenously expressing BDL103 under nitrogen limitingconditions BDL103 Long or Short/Event No. Long/ Long/ Long/ Long/ Long/Short/ Short/ Short/ Short/ Short/ 3054 3055 3056 3057 3058 3060 30613062 3063 3064 Roots P 0.34 0.03 Coverage A 1.10 1.26 (time point 6)Roots P 0.05 0.00 0.27 Coverage A 1.19 1.40 1.18 (time point 9) RootsLength P 0.04 (time point 1) A 1.11 Roots Length P 0.00 (time point 6) A1.20 Roots Length P 0.01 0.00 (time point 9) A 1.14 1.25 GR of Roots P0.21 0.03 Coverage A 1.19 1.31 (time point 6) GR of Roots P 0.53 0.090.37 0.01 0.14 0.34 Coverage A 1.19 1.37 1.27 1.70 1.66 1.26 (time point9) GR of Roots P 0.11 0.01 Length (time A 1.21 1.25 point 6) GR of RootsP 0.06 0.09 0.01 0.07 0.37 Length (time A 1.28 1.28 1.39 1.39 1.11 point9) RGR of P 0.08 0.00 0.21 0.06 0.04 0.00 Roots A 1.55 1.69 1.20 1.391.57 1.91 Coverage (time point 6) RGR of P 0.04 0.33 0.11 0.22 0.23 0.140.45 0.13 0.02 0.31 Roots A 2.06 1.23 1.95 1.43 1.32 1.52 1.23 1.71 2.531.50 Coverage (time point 9) RGR of P 0.07 0.01 0.32 0.31 0.12 0.05Roots Length A 1.34 1.39 1.11 1.14 1.24 1.39 (time point 6) RGR of P0.21 0.38 0.21 0.06 0.33 0.19 0.33 0.06 0.08 0.55 Roots Length A 1.351.15 1.27 1.32 1.13 1.29 1.22 1.40 1.67 1.22 (time point 9) DW [gr] P0.13 (time point 1) A 1.22 DW [gr] P 0.13 (time point 6) A 1.22 DW [gr]P 0.13 (time point 9) A 1.22 Table 59. Analysis of growth parameters intissue culture conditions of transgenic plants overexpressingBDL103-Short polynucleotide (SEQ ID NO: 671) or BDL103-Longpolynucleotide (SEQ ID NO: 670) under the regulation of a constitutivepromoter (35S; SEQ ID NO: 675) when grown under nitrogen limitingconditions (N 0.75 mM; see example 6). Each event number refers to anindependent transformation event in a plant (i.e., generation of atransgenic plant expressing the polynucleotide of choice). “A” =average; “P” = p-value; “RGR” = relative growth rate; “DW” = dry weight;“gr” = grams; Root coverage is presented in cm²; root length ispresented in cm; GR of root length is presented in cm/day; RGR of rootlength is presented in cm/day; RGR of root coverage is presented incm²/day. The various time points indicate days from beginning ofexperiment in which parameters were measured.

TABLE 60 Improved growth rate, root coverage, root length and biomass intransgenic plants exogenously expressing BDL103 under osmotic stressconditions BDL103 Long or Short/Event No. Long/ Long/ Long/ Long/ Long/Short/ Short/ Short/ Short/ Short/ 3054 3055 3056 3057 3058 3060 30613062 3063 3064 Roots P 0.08 0.08 Coverage A 1.25 1.34 (time point 9)Roots Length P 0.08 0.03 (time point 9) A 1.13 1.26 GR of Roots P 0.320.06 0.13 Coverage A 1.19 1.23 1.23 (time point 6) GR of Roots P 0.390.28 0.04 0.04 Coverage A 1.22 1.20 1.65 1.96 (time point 9) GR of RootsP 0.11 0.03 0.05 Length (time A 1.25 1.27 1.40 point 6) GR of Roots P0.09 0.02 0.01 0.02 Length (time A 1.23 1.28 1.45 1.80 point 9) RGR of P0.00 0.01 0.05 0.68 Roots A 2.12 1.84 2.14 1.16 Coverage (time point 6)RGR of P 0.46 0.04 0.27 0.24 0.56 0.02 0.01 0.63 Roots A 1.26 1.42 1.201.67 1.24 1.50 1.80 1.26 Coverage (time point 9) RGR of P 0.00 0.00 0.05Roots Length A 1.64 1.57 1.76 (time point 6) RGR of P 0.18 0.09 0.030.57 0.15 0.01 0.02 Roots Length A 1.26 1.28 1.21 1.23 1.32 1.40 1.65(time point 9) DW [gr] P 0.04 0.05 0.41 (time point 1) A 1.41 1.30 1.22DW [gr] P 0.04 0.05 0.41 (time point 6) A 1.41 1.30 1.22 DW [gr] P 0.040.05 0.41 (time point 9) A 1.41 1.30 1.22 FW [gr] P 0.47 0.17 0.30 (timepoint 1) A 1.32 1.23 1.36 FW [gr] P 0.47 0.17 0.30 (time point 6) A 1.321.23 1.36 FW [gr] P 0.47 0.17 0.30 (time point 9) A 1.32 1.23 1.36 Table60. Analysis of growth parameters in tissue culture conditions oftransgenic plants overexpressing BDL103-Short polynucleotide (SEQ ID NO:671) or BDL103-Long polynucleotide (SEQ ID NO: 670) under the regulationof a constitutive promoter (35S; SEQ ID NO: 675) when grown underosmotic stress condition in the presence of 15% PEG (polyethyleneglycol). Each event number refers to an independent transformation eventin a plant (i.e., generation of a transgenic plant expressing thepolynucleotide of choice). “A” = average; “P” = p-value; “GR” = growthrate; “RGR” = relative growth rate; “DW” = dry weight; “FW” = freshweigh; “gr” = grams; Root coverage is presented in cm²; root length ispresented in cm; GR of root length is presented in cm/day; RGR of rootlength is presented in cm/day; RGR of root coverage is presented incm²/day. The various time points indicate days from beginning ofexperiment in which parameters were measured.

Example 11 Transgenic Arabidopsis Plants which Exogenously ExpressBDL103 Exhibit Increased Commercially Desired Traits in a GreenhouseAssay

Greenhouse assays were performed as described in Example 7 hereinabovefor determining plant performance under normal conditions (i.e.,irrigation with tap water).

Tables 61-62 depict analyses of growth rate, biomass, rosette diameter,rosette area, plot coverage, leaf number, petiole relative area, leafblade area, blade relative area and harvest index in plantsoverexpressing the BDL103-long (SEQ ID NO:670; Table 61) and theBDL103-short (SEQ ID NO:671; Table 62) polynucleotides under theregulation of a constitutive (35S; SEQ ID NO:675) when grown in agreenhouse under normal conditions until seed production. Each Tableincludes data of several transformation events per gene. Results wereconsidered significant if p-value was lower than 0.1 when compared tocontrol plants (transformed with an empty vector).

TABLE 61 Improved growth rate, biomass, rosette diameter, rosette area,plot coverage, leaf number, petiole relative area, leaf blade area,blade relative area and harvest index in transgenic arabidopsis plantsexogenously expressing BDL103-long (SEQ ID NO: 670) under favorableconditions Event No. Parameter 2541 2541 2542 2542 2543 2543 2545 25452546 2546 (time point) A P A P A P A P A P Yield 1.76 0.40 1.18 0.431.51 0.05 1.36 0.03 Rosette Diameter 1.11 0.18 1.37 0.03 1.33 0.10 1.140.41 1.12 0.65 (time point 8) Rosette Diameter 1.13 0.00 1.31 0.22 1.250.10 1.11 0.43 1.13 (time point 5) Rosette Diameter 1.59 0.01 1.52 0.071.29 0.25 1.16 0.55 (time point 3) Rosette Diameter 1.20 0.08 1.19 0.30(time point 1) Rosette Area 1.37 0.00 1.65 0.24 1.53 0.14 1.24 0.35(time point 8) Rosette Area 1.23 0.12 1.48 0.17 1.25 0.19 1.21 (timepoint 5) Rosette Area 1.97 0.16 1.68 0.06 1.37 0.38 1.23 0.60 (timepoint 3) Rosette Area 1.17 0.36 1.23 0.04 (time point 1) RGR of Rosette1.13 0.54 1.14 0.07 1.38 Diameter (time point 8) RGR of Rosette 9.610.11 Diameter (time point 5) RGR of Rosette 2.20 0.18 2.00 0.11 2.000.11 2.41 0.04 Diameter (time point 3) RGR of Rosette Area 1.18 0.361.15 0.30 1.32 0.00 1.19 0.00 1.19 (time point 8) RGR of Rosette Area6.59 0.29 (time point 5) RGR of Rosette Area 2.22 0.28 2.66 0.00 2.670.26 2.11 0.12 (time point 3) RGR of Plot 1.18 0.36 1.15 0.30 1.32 0.001.19 0.00 1.19 Coverage (time point 8) RGR of Plot 6.59 0.29 Coverage(time point 5) RGR of Plot 2.22 0.28 2.66 0.00 2.67 0.26 2.11 0.12Coverage (time point 3) RGR of Leaf Number 8.30 0.21 (time point 5) RGRof Leaf Number 1.39 0.44 2.59 0.01 3.00 0.33 2.37 0.02 (time point 3)Plot Coverage 1.40 0.00 1.59 0.34 1.55 0.13 1.26 0.32 (time point 8)Plot Coverage 1.24 0.10 1.42 0.32 1.27 0.17 1.11 0.53 (time point 5)Plot Coverage 1.90 0.26 1.71 0.05 1.39 0.36 1.15 0.64 (time point 3)Plot Coverage 1.19 0.33 1.17 0.09 (time point 1) Petiole Relative Area1.31 0.81 1.23 0.66 (time point 8) Petiole Relative Area 2.69 0.02 (timepoint 3) Petiole Relative Area 1.18 0.43 1.42 0.00 1.39 0.26 (timepoint 1) Leaf Petiole Area 1.95 0.64 1.67 0.22 (time point 8) LeafPetiole Area 3.05 0.00 (time point 3) Leaf Petiole Area 1.11 0.61 1.330.01 1.18 0.49 (time point 1) Leaf Number 1.10 0.02 1.10 0.03 (timepoint 8) Leaf Number 1.07 0.05 1.12 0.15 1.11 0.28 1.12 (time point 5)Leaf Number 1.50 0.02 1.43 0.12 1.25 0.40 1.28 0.29 (time point 3) LeafNumber 1.28 0.00 1.37 0.00 (time point 1) Leaf Blade Area 1.43 0.00 1.530.32 1.37 0.15 1.28 0.00 (time point 8) Leaf Blade Area 1.18 0.17 1.360.14 1.14 0.11 1.11 (time point 5) Leaf Blade Area 1.59 0.20 1.41 0.021.31 0.14 1.14 0.59 (time point 3) Harvest index 1.19 0.01 1.18 0.381.13 0.08 Blade Relative Area 1.03 0.07 (time point 8) Blade RelativeArea 1.03 0.05 (time point 5) Blade Relative Area 1.19 0.08 1.18 0.091.18 0.10 1.18 0.09 (time point 3) Blade Relative Area 1.11 0.01 (timepoint 1) Table 61. Analysis of growth parameters in a greenhouse assayof transgenic plants overexpressing BDL103-long polynucleotide (SEQ IDNO: 670) under the regulation of a constitutive promoter (35S; SEQ IDNO: 675) which were grown until seed production under normal conditions(as described in Example 7 above). Each event number refers to anindependent transformation event in a plant (i.e., generation of atransgenic plant expressing the polynucleotide of choice). “A” =average; “P” = p-value; “RGR” = relative growth rate; “gr” = grams;yield is presented in mg/plant; Rosette Diameter is presented incm/plant; Rosette Area is presented in cm²/plant; RGR of RosetteDiameter is presented in cm/plant*day; RGR of Rosette Area is presentedin cm²/plant*day; RGR of Plot Coverage is presented in cm²/plant*day;RGR of Leaf Number is presented in 1/day; Plot Coverage is presented incm²; Petiole Relative Area is presented in percent; Leaf Petiole Area ispresented in cm²; Leaf Number is presented as number of leaves perplant; Leaf Blade Area is presented in cm²; Harvest Index is presentedin g/DW (dry weight); Blade Relative Area is presented in percent; Thevarious time points indicate days from beginning of experiment in whichparameters were measured.

TABLE 62 Improved growth rate, biomass, rosette diameter, rosette area,plot coverage, leaf number, petiole relative area, leaf blade area,blade relative area and harvest index in transgenic arabidopsis plantsexogenously expressing BDL103-short (SEQ ID NO: 671) under favorableconditions Event No. Parameter 2353 2353 2357 2357 2359 2359 2360 23602361 2361 (time point) A P A P A P A P A P Yield 1.32 0.26 1.32 0.011.14 0.18 Rosette Diameter 1.28 1.24 0.00 1.13 0.44 1.21 0.31 1.15 0.05(time point 8) Rosette Diameter 1.26 0.07 1.15 0.13 (time point 5)Rosette Diameter 1.43 0.01 1.28 0.06 1.21 0.13 1.29 0.41 1.10 0.52 (timepoint 3) Rosette Diameter 1.26 0.29 (time point 1) Rosette Area 1.461.36 0.00 1.12 0.10 1.32 0.39 1.15 0.23 (time point 8) Rosette Area 1.320.00 1.27 0.00 1.16 0.61 (time point 5) Rosette Area 1.36 0.10 1.56 0.021.31 0.63 (time point 3) Rosette Area 1.18 0.06 (time point 1) RGR ofRosette 1.13 1.19 0.32 1.26 0.52 1.29 0.22 1.37 0.14 Diameter (timepoint 8) RGR of Rosette 1.52 0.46 2.09 0.08 1.94 0.12 1.64 0.42 1.660.46 Diameter (time point 3) RGR of Rosette Area 1.15 1.18 0.07 1.210.03 1.23 0.01 (time point 8) RGR of Rosette Area 1.34 0.34 2.01 0.041.58 0.34 1.77 0.11 2.15 0.25 (time point 3) RGR of Plot 1.15 1.18 0.071.21 0.03 1.23 0.01 Coverage (time point 8) RGR of Plot 1.34 0.34 2.010.04 1.58 0.34 1.77 0.11 2.15 0.25 Coverage (time point 3) RGR of LeafNumber 1.19 (time point 8) RGR of Leaf Number 1.22 0.63 1.42 0.56 2.510.13 1.75 0.25 3.70 0.00 (time point 3) Plot Coverage 1.48 1.29 0.101.35 0.37 1.17 0.19 (time point 8) Plot Coverage 1.34 0.00 1.21 0.061.18 0.58 (time point 5) Plot Coverage 1.39 0.09 1.49 0.06 1.33 0.611.11 0.75 (time point 3) Plot Coverage 1.20 0.06 (time point 1) PetioleRelative Area 1.21 0.71 1.92 0.25 3.37 0.44 (time point 8) PetioleRelative Area 1.20 0.25 (time point 5) Petiole Relative Area 1.31 0.201.75 0.01 (time point 1) Leaf Petiole Area 1.32 0.60 2.47 0.32 3.96 0.43(time point 8) Leaf Petiole Area 1.20 0.38 (time point 5) Leaf PetioleArea 1.24 0.02 1.96 0.21 (time point 1) Leaf Number 1.15 (time point 8)Leaf Number 1.12 0.01 1.21 0.00 1.08 0.05 (time point 5) Leaf Number1.31 0.09 1.32 0.11 1.19 0.29 1.28 0.45 1.15 0.63 (time point 3) LeafNumber 1.26 0.01 1.21 0.23 (time point 1) Leaf Blade Area 1.29 1.33 0.051.08 0.05 1.22 0.39 (time point 8) Leaf Blade Area 1.21 0.01 1.10 0.031.11 0.63 (time point 5) Leaf Blade Area 1.22 0.16 1.42 0.02 (time point3) Harvest index 1.30 0.44 Blade Relative Area 1.04 0.07 (time point 5)Blade Relative Area 1.15 0.16 1.19 0.09 1.10 0.28 (time point 3) Table62. Analysis of growth parameters in a greenhouse assay of transgenicplants overexpressing BDL103-short polynucleotide (SEQ ID NO: 671) underthe regulation of a constitutive promoter (35S; SEQ ID NO: 675) whichwere grown until seed production under normal conditions (as describedin Example 7 above). Each event number refers to an independenttransformation event in a plant (i.e., generation of a transgenic plantexpressing the polynucleotide of choice). “A” = average; “P” = p-value;“RGR” = relative growth rate; “gr” = grams; yield is presented inmg/plant; Rosette Diameter is presented in cm/plant; Rosette Area ispresented in cm²/plant; RGR of Rosette Diameter is presented incm/plant*day; RGR of Rosette Area is presented in cm²/plant*day; RGR ofPlot Coverage is presented in cm²/plant*day; RGR of Leaf Number ispresented in 1/day; Plot Coverage is presented in cm²; Petiole RelativeArea is presented in percent; Leaf Petiole Area is presented in cm²;Leaf Number is presented as number of leaves per plant; Leaf Blade Areais presented in cm²; Harvest Index is presented in g/DW (dry weight);Blade Relative Area is presented in percent; The various time pointsindicate days from beginning of experiment in which parameters weremeasured.

Although the invention has been described in conjunction with specificembodiments thereof, it is evident that many alternatives, modificationsand variations will be apparent to those skilled in the art.Accordingly, it is intended to embrace all such alternatives,modifications and variations that fall within the spirit and broad scopeof the appended claims.

All publications, patents and patent applications mentioned in thisspecification are herein incorporated in their entirety by referenceinto the specification, to the same extent as if each individualpublication, patent or patent application was specifically andindividually indicated to be incorporated herein by reference. Inaddition, citation or identification of any reference in thisapplication shall not be construed as an admission that such referenceis available as prior art to the present invention. To the extent thatsection headings are used, they should not be construed as necessarilylimiting.

What is claimed is:
 1. A method of increasing abiotic stress tolerance,yield, biomass, growth rate, vigor, oil content, fiber yield, fiberquality, and/or nitrogen use efficiency of a plant, comprisingexpressing within the plant an exogenous polynucleotide comprising anucleic acid sequence encoding a polypeptide which exhibits at least 80%sequence identity to SEQ ID NO: 82, 73, 652, 71, 86, 60-70, 72, 74,76-85, 87-95, 108-109, 112, 359-589, 602-604, 653-660, 665, 668, or 672,thereby increasing the abiotic stress tolerance, yield, biomass, growthrate, vigor, oil content, fiber yield, fiber quality, and/or nitrogenuse efficiency of the plant.
 2. The method of claim 1, wherein saidpolypeptide exhibits at least 90% sequence identity to the polypeptideselected from the group consisting of SEQ ID NOs: 82, 73, 652, 71, 86,60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589, 591-597,600-604, 653-662, 664, 666-669, and
 672. 3. The method of claim 1,wherein said polypeptide exhibits at least 95% sequence identity to thepolypeptide selected from the group consisting of SEQ ID NOs: 82, 73,652, 71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589,591-597, 600-604, 653-662, 664, 666-669, and
 672. 4. The method of claim1, wherein said polypeptide is selected from the group consisting of SEQID NOs: 82, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109, 111,112, 359-589, 591-597, 600-604, 653-662, 664, 666-669, and
 672. 5. Themethod of claim 1, wherein said nucleic acid sequence encoding saidpolypeptide exhibits at least 80% sequence identity to SEQ ID NO: 625,23, 617, 606, 615, 629, 1-15, 17-36, 40, 41, 43-45, 49, 52-56, 58,113-343, 351, 354-358, 605, 607-614, 616, 618, 620-628, 630-638, 642,645, 650, 651, 670, or
 671. 6. The method of claim 1, wherein saidnucleic acid sequence encoding said polypeptide is selected from thegroup consisting of SEQ ID NOs: 625, 23, 617, 606, 615, 629, 1-15,17-49, 51-59, 113-343, 345-351, 353-358, 605, 607-614, 616, 618,620-628, 630-638, 641, 642, 644, 644-646, 648-651, 670, and
 671. 7. Anisolated polynucleotide comprising a nucleic acid sequence encoding apolypeptide comprising an amino acid sequence which exhibits at least80% sequence identity to the amino acid sequence set forth in SEQ ID NO:82, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-95, 108-109, 112, 359-589,602-604, 653-660, 665, 668, or 672, wherein said amino acid sequence iscapable of increasing abiotic stress tolerance, yield, biomass, growthrate, vigor, oil content, fiber yield, fiber quality, and/or nitrogenuse efficiency of a plant.
 8. The isolated polynucleotide of claim 7,wherein said amino acid sequence exhibits at least 90% sequence identityto the amino acid sequence selected from the group consisting of SEQ IDNOs: 82, 73, 652, 71, 86, 60-70, 72, 74, 76-85, 87-98, 100-109, 111,112, 359-589, 591-597, 600-604, 653-662, 664, 666-669, and
 672. 9. Theisolated polynucleotide of claim 7, wherein said amino acid sequenceexhibits at least 95% sequence identity to the amino acid sequenceselected from the group consisting of SEQ ID NOs: 82, 73, 652, 71, 86,60-70, 72, 74, 76-85, 87-98, 100-109, 111, 112, 359-589, 591-597,600-604, 653-662, 664, 666-669, and
 672. 10. The isolated polynucleotideof claim 7, wherein said nucleic acid sequence encoding said polypeptideexhibits at least 80% sequence identity to SEQ ID NO: 625, 23, 617, 606,615, 629, 1-15, 17-36, 40, 41, 43-45, 49, 52-56, 58, 113-343, 351,354-358, 605, 607-614, 616, 618, 620-628, 630-638, 642, 645, 650-651,670, or
 671. 11. The isolated polynucleotide of claim 7, wherein saidnucleic acid sequence is selected from the group consisting of SEQ IDNOs: 625, 23, 617, 606, 615, 629, 1-15, 17-49, 51-59, 113-343, 345-351,353-358, 605, 607-614, 616, 618, 620-628, 630-638, 641, 642, 644,644-646, 648-651, 670, and
 671. 12. A nucleic acid construct comprisingthe isolated polynucleotide of claim 7, and a promoter for directingtranscription of said nucleic acid sequence in a host cell.
 13. A plantcell transformed with the nucleic acid construct of claim
 12. 14. Theplant cell of claim 13, wherein said plant cell forms part of a plant.15. A transgenic plant comprising the nucleic acid construct of claim12.
 16. The method of claim 1, further comprising growing the plantexpressing said exogenous polynucleotide under the abiotic stress. 17.The method of claim 1, wherein said abiotic stress is selected from thegroup consisting of salinity, drought, water deprivation, flood,etiolation, low temperature, high temperature, heavy metal toxicity,anaerobiosis, nutrient deficiency, nutrient excess, atmosphericpollution and UV irradiation.
 18. The method of claim 1, furthercomprising selecting the plant expressing said exogenous polynucleotidefor an increased abiotic stress tolerance, yield, biomass, growth rate,oil content, fiber yield, fiber quality, and/or nitrogen use efficiencyas compared to a non-transformed plant which is grown under the samegrowth conditions.
 19. The method of claim 18, wherein said selecting isperformed under non-stress conditions.
 20. A method of producing seedsof a crop, comprising: (a) selecting a parent plant being transformedwith an exogenous polynucleotide encoding a polypeptide comprising anamino acid sequence which exhibits at least 80% sequence identity to thepolypeptide selected from the group consisting of SEQ ID NOs: 82, 73,652, 71, 86, 60-70, 72, 74, 76-85, 87-95, 108-109, 112, 359-589,602-604, 653-660, 665, 668, and 672, said parent plant exhibits anincreased trait selected from the group consisting of: increased abioticstress tolerance, increased yield, increased biomass, increased growthrate, increased oil content, increased fiber yield, increased fiberquality, or increased nitrogen use efficiency as compared to anon-transformed plant which is grown under the same growth conditions,and; (b) growing a seed producing plant from said parent plant resultantof step (a), wherein said seed producing plant which comprises saidexogenous polynucleotide has said increased trait, and; (c) producingseeds from said seed producing plant resultant of step (b), therebyproducing seeds of the crop.